Metalloenzyme inhibitor compounds

ABSTRACT

The instant invention describes compounds having metalloenzyme modulating activity, and methods of treating diseases, disorders or symptoms thereof mediated by such metalloenzymes.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.16/259,096, filed Jan. 28, 2019, the entire content of which is herebyincorporated herein by reference. U.S. application Ser. No. 16/259,096is a continuation of U.S. application Ser. No. 14/373,314, filed Jul.18, 2014, the entire contents of which is incorporated herein byreference. U.S. application Ser. No. 14/373,314, is a national phase ofInternational Application No. PCT/US2013/022313, filed Jan. 18, 2013,the entire contents of which is incorporated herein by reference. U.S.application Ser. No. 14/373,314 also claims the benefit of U.S.Provisional Application No. 61/589,076 filed Jan. 20, 2012, the contentsof which is incorporated herein by reference in its entirety.

BACKGROUND

Living organisms have developed tightly regulated processes thatspecifically imports metals, transport them to intracellular storagesites and ultimately transport them to sites of use. One of the mostimportant functions of metals such as zinc and iron in biologicalsystems is to enable the activity of metalloenzymes. Metalloenzymes areenzymes that incorporate metal ions into the enzyme active site andutilize the metal as a part of the catalytic process. More thanone-third of all characterized enzymes are metalloenzymes.

The function of metalloenzymes is highly dependent on the presence ofthe metal ion in the active site of the enzyme. It is well recognizedthat agents which bind to and inactivate the active site metal iondramatically decrease the activity of the enzyme. Nature employs thissame strategy to decrease the activity of certain metalloenzymes duringperiods in which the enzymatic activity is undesirable. For example, theprotein TIMP (tissue inhibitor of metalloproteases) binds to the zincion in the active site of various matrix metalloprotease enzymes andthereby arrests the enzymatic activity. The pharmaceutical industry hasused the same strategy in the design of therapeutic agents. For example,the azole antifungal agents fluconazole and voriconazole contain a1-(1,2,4-triazole) group that binds to the heme iron present in theactive site of the target enzyme lanosterol demethylase and therebyinactivates the enzyme. Another example includes the zinc-bindinghydroxamic acid group that has been incorporated into most publishedinhibitors of matrix metalloproteinases and histone deacetylases.Another example is the zinc-binding carboxylic acid group that has beenincorporated into most published angiotensin-converting enzymeinhibitors.

In the design of clinically safe and effective metalloenzyme inhibitors,use of the most appropriate metal-binding group for the particulartarget and clinical indication is critical. If a weakly bindingmetal-binding group is utilized, potency may be suboptimal. On the otherhand, if a very tightly binding metal-binding group is utilized,selectivity for the target enzyme versus related metalloenzymes may besuboptimal. The lack of optimal selectivity can be a cause for clinicaltoxicity due to unintended inhibition of these off-targetmetalloenzymes. One example of such clinical toxicity is the unintendedinhibition of human drug metabolizing enzymes such as CYP2C9, CYP2C19and CYP3A4 by the currently-available azole antifungal agents such asfluconazole and voriconazole. It is believed that this off-targetinhibition is caused primarily by the indiscriminate binding of thecurrently utilized 1-(1,2,4-triazole) to iron in the active site ofCYP2C9, CYP2C19 and CYP3A4. Another example of this is the joint painthat has been observed in many clinical trials of matrixmetalloproteinase inhibitors.

This toxicity is considered to be related to inhibition of off-targetmetalloenzymes due to indiscriminate binding of the hydroxamic acidgroup to zinc in the off-target active sites.

Therefore, the search for metal-binding groups that can achieve a betterbalance of potency and selectivity remains an important goal and wouldbe significant in the realization of therapeutic agents and methods toaddress currently unmet needs in treating and preventing diseases,disorders and symptoms thereof.

BRIEF SUMMARY OF THE INVENTION

The invention is directed towards compounds (e.g., any of thosedelineated herein), methods of modulating activity of metalloenzymes,and methods of treating diseases, disorders or symptoms thereof. Themethods can comprise the compounds herein.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is H, halo, alkyl or haloalkyl;

R₂ is H, halo, alkyl or haloalkyl;

R₃ is independently H, alkyl, cyano, haloalkyl, alkoxy, halo,haloalkoxy, cycloalkyl, alkoxyalkyl, haloalkoxyalkyl, aryloxyalkyl,thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇, nitro,C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino (such as morpholino,pyrrolidino, piperidino, N-alkyl piperidino);

R₄ is heteroaryl or cycloalkyl, optionally substituted with 0, 1, 2 or 3independent R₃;

R₅ is H, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

R₆ is H, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is alkyl or cycloalkyl;

R₈ is alkyl or haloalkyl; and

n is 0, 1, 2 or 3.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is H, halo, alkyl or haloalkyl;

R₂ is H, halo, alkyl or haloalkyl;

R₃ is independently H, alkyl, cyano, haloalkyl, alkoxy, halo,haloalkoxy, cycloalkyl, alkoxyalkyl, haloalkoxyalkyl, aryloxyalkyl,thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇, nitro,C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino (such as morpholino,pyrrolidino, piperidino, N-alkyl piperidino);

R₄ is aryl, heteroaryl or cycloalkyl, optionally substituted with 0, 1,2 or 3 independent R₃;

R₅ is H, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

R₆ is H, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is alkyl or cycloalkyl;

R₈ is alkyl or haloalkyl; and

n is 0, 1, 2 or 3.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is H, halo, alkyl or haloalkyl;

R₂ is H, halo, alkyl or haloalkyl;

R₃ is independently H, alkyl, cyano, haloalkyl, alkoxy, halo,haloalkoxy, cycloalkyl, alkoxyalkyl, haloalkoxyalkyl, aryloxyalkyl,thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇, nitro,C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino, NHC(═O)CF₃,OCF₂C(═O)OR₇, (such as morpholino, pyrrolidino, piperidino, N-alkylpiperidino);

R₄ is aryl, heteroaryl or cycloalkyl, optionally substituted with 0, 1,2 or 3 independent R₃;

R₅ is H, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

R₆ is H, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is alkyl or cycloalkyl;

R₈ is alkyl or haloalkyl; and

n is 0, 1, 2 or 3.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is H, halo, alkyl or haloalkyl;

R₂ is H, halo, alkyl or haloalkyl;

R₃ is independently cycloalkyl, alkoxyalkyl, haloalkoxyalkyl,aryloxyalkyl, thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇,nitro, C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino (such asmorpholino, pyrrolidino, piperidino, N-alkyl piperidino);

R₄ is heteroaryl or cycloalkyl, optionally substituted with 0, 1, 2 or 3independent R₃;

R₅ is H, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

R₆ is H, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is alkyl or cycloalkyl;

R₈ is alkyl or haloalkyl; and

n is 0, 1, 2 or 3.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is halo;

preferably R₁ is fluoro;

R₂ is halo;

preferably R₂ is fluoro;

preferably R₁ and R₂ are fluoro;

R₃ is independently independently H, alkyl, cyano, haloalkyl, alkoxy,halo, haloalkoxy, cycloalkyl, alkoxyalkyl, haloalkoxyalkyl,aryloxyalkyl, thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇,nitro, C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino, NHC(═O)CF₃,or OCF₂C(═O)OR₇;

R₄ is aryl, heteroaryl or cycloalkyl, optionally substituted with 0, 1,2 or 3 independent R₃;

R₅ is —P(O)(OH)₂ or —CH₂—O—P(O)(OH)₂;

R₆ is hydrogen, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is hydrogen, alkyl or cycloalkyl;

R₈ is hydrogen, alkyl or haloalkyl;

n is 0, 1, 2 or 3; and

preferably n is 1, 2, or 3.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is halo;

preferably R₁ is fluoro;

R₂ is halo;

preferably R₂ is fluoro;

preferably R₁ and R₂ are fluoro;

R₃ is independently cycloalkyl, alkoxyalkyl, haloalkoxyalkyl,aryloxyalkyl, thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇,nitro, C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino, NHC(═O)CF₃,or OCF₂C(═O)OR₇;

R₄ is aryl, heteroaryl or cycloalkyl, optionally substituted with 0, 1,2 or 3 independent R₃;

R₅ is H, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

preferably R₅ is H;

R₆ is hydrogen, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is hydrogen, alkyl or cycloalkyl;

R₈ is hydrogen, alkyl or haloalkyl;

n is 0, 1, 2 or 3; and

preferably n is 1, 2, or 3.

A compound of Formula I, or salt, solvate, hydrate or prodrug thereof,wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, or optionally substituted pyrazolyl;

R₁ is H, halo, alkyl or haloalkyl;

preferably R₁ is alkyl;

preferably R₁ is methyl;

R₂ is H, alkyl or haloalkyl;

R₃ is independently H, alkyl, cyano, haloalkyl, alkoxy, halo,haloalkoxy, cycloalkyl, alkoxyalkyl, haloalkoxyalkyl, aryloxyalkyl,thioalkyl, hydroxyl, halothioalkyl, thiocyanate, S(O)₂R₇, nitro,C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic amino, NHC(═O)CF₃, orOCF₂C(═O)OR₇;

R₄ is aryl, heteroaryl, or cycloalkyl, optionally substituted with 0, 1,2 or 3 independent R₃;

R₅ is H, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

preferably R₅ is H;

preferably R₅ is —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyl optionallysubstituted with amino;

R₆ is H, halo, alkyl, haloalkyl or haloalkoxy;

R₇ is H, alkyl or cycloalkyl;

R₈ is H, alkyl or haloalkyl;

n is 0, 1, 2 or 3; and

preferably n is 1, 2, or 3.

Another aspect is a compound of the formulae herein, wherein thecompound is not4-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)phenol(23) or2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-((trifluoromethyl)thio)phenyl)pyridin-2-yl)propan-2-ol(27).

Another aspect is a compound of the formulae herein, wherein the MBG isan optionally substituted 1H-tetrazol-1-yl, optionally substituted2H-tetrazol-2-yl, optionally substituted 1H-1,2,4-triazol-1-yl,optionally substituted 1H-1,2,3-triazol-1-yl, or optionally substituted1H-pyrazol-3-yl.

Another aspect is a compound of the formulae herein, wherein the MBG isunsubstituted 1H-tetrazol-1-yl, unsubstituted 2H-tetrazol-2-yl,unsubstituted 1H-1,2,4-triazol-1-yl, unsubstituted1H-1,2,3-triazol-1-yl, or unsubstituted 1H-pyrazol-3-yl.

Another aspect is a compound of the formulae herein, wherein R₁ isfluoro.

Another aspect is a compound of the formulae herein, wherein R₂ isfluoro.

Another aspect is a compound of the formulae herein, wherein R₁ and R₂are fluoro.

Another aspect is a compound of the formulae herein, wherein R₁ isalkyl.

Another aspect is a compound of the formulae herein, wherein R₁ ismethyl.

Another aspect is a compound of the formulae herein, wherein R₁ ismethyl and R₂ is fluoro.

Another aspect is a compound of the formulae herein, wherein R₃ isindependently cycloalkyl, alkoxyalkyl, haloalkoxyalkyl, aryloxyalkyl,thioalkyl, thiocyanate, S(O)₂R₇, nitro, C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈,amino, cyclic amino.

Another aspect is a compound of the formulae herein, wherein R₅ is H.

Another aspect is a compound of the formulae herein, wherein R₅ is aminosubstituted acyl.

Another aspect is a compound of the formulae herein, wherein R₆ is H.

Another aspect is a compound of the formulae herein, wherein R₆ is halo,alkyl, haloalkyl or haloalkoxy.

Another aspect is a compound of the formulae herein, wherein n is 1, 2or 3.

Another aspect is a compound of the formulae herein, wherein:

-   -   R₁ is fluoro;    -   R₂ is fluoro; and    -   R₅ is H.

Another aspect is a compound of the formulae herein, wherein:

-   -   R₁ is methyl;    -   R₂ is fluoro; and    -   R₃ is independently H, alkyl, cyano, haloalkyl, alkoxy, halo,        haloalkoxy, cycloalkyl, alkoxyalkyl, haloalkoxyalkyl,        aryloxyalkyl, thioalkyl, hydroxyl, halothioalkyl, thiocyanate,        S(O)₂R₇, nitro, C(═O)CF₃, C(═O)OR₇, C(═O)NR₇R₈, amino, cyclic        amino (such as morpholino, pyrrolidino, piperidino, N-alkyl        piperidino).

Another aspect is a compound of the formulae herein, wherein:

-   -   n is 1 or 2.

Another aspect is a compound of the formulae herein, wherein:

-   -   n is 1.

Another aspect is a compound of the formulae herein, wherein:

-   -   each R₃ is independently 4-cyano, 4-trifluoromethyl, 3-cyano,        4-isopropoxy, 4-fluoro, 3-trifluoromethoxy, 4-trifluoromethoxy,        3-chloro, 4-chloro, 2-fluoro, 5-fluoro,        4-(2,2,2-trifluoroethoxy), 4-(3,3,3-trifluoro,        2,2-difluoropropoxy)), 2,5-difluoro, 3-fluoro, 4-hydroxy,        3-isopropyl, 3,4-difluoro, 3-difluoromethoxy,        4-trifluoromethylthio, 4-t-butoxy, 4-chloro-3-fluoro, 3-hydroxy,        3-trifluoromethyl, 4-nitro, 4-trifluoromethylcarbonyl, H,        4-morpholino, 4-(trifluoroacetamido), 4-(difluoromethoxy),        4-(difluoromethylthio), 4-(2,2,2-trifluoroethyl),        4-(methylamido), 4-(—O—CF₂C(O)OEt), 4-(3,3,3-trifluoropropoxy),        or 4-(2,2,2-trifluoroethylthio).

In one aspect, the compound of Formula I is that wherein the compoundinhibits (or is identified to inhibit) lanosterol demethylase (CYP51).

In one aspect, the compound of Formula I is that wherein the compound isidentified as having an activity range against a target organism orenzyme and an activity range against an off-target enzyme (e.g., C.albicans MIC<0.02 μg/mL and IC₅₀>16 μM for CYP2C9, CYP2C19 and CYP3A4;C. albicans MIC<0.10 μg/mL and IC₅₀>10 μM for CYP2C9, CYP2C19 andCYP3A4; C. albicans MIC<0.5 μg/mL and IC₅₀>15 μM for CYP2C9, CYP2C19 andCYP3A4).

The compounds herein include those wherein the compound is identified asattaining affinity, at least in part, for a metalloenzyme by formationof one or more of the following types of chemical interactions or bondsto a metal:sigma bonds, covalent bonds, coordinate-covalent bonds, ionicbonds, pi bonds, delta bonds, or backbonding interactions. The compoundscan also attain affinity through a weaker interaction with the metalsuch as van der Waals interactions, pi-cation interactions, pi-anioninteractions, dipole-dipole interactions, ion-dipole interactions. Inone aspect, the compound is identified as having a bonding interactionwith the metal via the 1-tetrazolyl moiety; in another aspect, thecompound is identified as having a bonding interaction with the metalvia the N2 of the 1-tetrazolyl moiety; in another aspect, the compoundis identified as having a bonding interaction with the metal via the N3of the 1-tetrazolyl moiety; in another aspect, the compound isidentified as having a bonding interaction with the metal via the N4 ofthe 1-tetrazolyl moiety.

Methods for assessing metal-ligand binding interactions are known in theart as exemplified in references including, for example, “Principles ofBioinorganic Chemistry” by Lippard and Berg, University Science Books,(1994); “Mechanisms of Inorganic Reactions” by Basolo and Pearson JohnWiley & Sons Inc; 2nd edition (September 1967); “Biological InorganicChemistry” by Ivano Bertini, Harry Gray, Ed Stiefel, Joan Valentine,University Science Books (2007); Xue et al. “Nature Chemical Biology”,vol. 4, no. 2, 107-109 (2008).

In certain instances, the compounds of the invention are selected fromthe following of Formula I (and pharmaceutically acceptable salts,solvates, or hydrates thereof)

-   1-(5-(4-(tert-Butoxy)phenyOpyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (28);-   1-(5-(4-Chloro-3-fluorophenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (29);-   3-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)phenol    (30);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(3-(trifluoromethyl)phenyOpyridin-2-yl)propan-2-ol    (31);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-nitrophenyl)pyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (32);-   1-(4-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)phenyl)-2,2,2-trifluoroethanone    (33);-   2-(4-Chloro-2-fluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyridin-2-yl)propan-2-ol    (34);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-phenylpyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (35);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-morpholinophenyl)pyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (36);-   N-(4-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)phenyl)-2,2,2-trifluoroacetamide    (37);-   1-(5-(4-(Difluoromethoxy)phenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (38);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol    (39);-   1-(5-(4-((Difluoromethyl)thio)phenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (40);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethyl)phenyOpyridin-2-yl)propan-2-ol    (41);-   4-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)-N-methylbenzamide    (42);-   Ethyl    2-(4-(6-(2-(2,4-difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)phenoxy)-2,2-difluoroacetate    (43);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(3,3,3-trifluoropropoxy)phenyl)pyridin-2-yl)propan-2-ol    (44);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-((2,2,2-trifluoroethyl)thio)phenyl)pyridin-2-yl)propan-2-ol    (45);-   2-(2,4-Difluorophenyl)-3-fluoro-1-(1H-tetrazol-1-yl)-3-(5-(4-(trifluoromethoxy)phenyl)pyridin-2-yl)butan-2-ol    (46 and 47);-   2-(2,4-Difluorophenyl)-3-fluoro-3-(5-(4-fluorophenyl)pyridin-2-yl)-1-(2H-tetrazol-2-yl)butan-2-ol    (48 and 49);-   2-(2,4-Difluorophenyl)-3-fluoro-3-(5-(4-fluorophenyl)pyridin-2-yl)-1-(1H-tetrazol-1-yl)butan-2-ol    (50 and 51);-   2-(2,4-Difluorophenyl)-3-fluoro-1-(1H-tetrazol-1-yl)-3-(5-(4-(trifluoromethyl)phenyOpyridin-2-yl)butan-2-ol    (52 and 53);-   3-(5-(4-Chlorophenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-3-fluoro-1-(1H-tetrazol-1-yl)butan-2-ol    (54 and 55);-   2-(2,4-Difluorophenyl)-3-fluoro-1-(1H-tetrazol-1-yl)-3-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)butan-2-ol    (56 and 57);-   2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl    dihydrogen phosphate (58).

In another aspect, the invention provides a pharmaceutical compositioncomprising the compound of Formula I and a pharmaceutically acceptablecarrier.

In other aspects, the invention provides a method of modulatingmetalloenzyme activity in a subject, comprising contacting the subjectwith a compound of Formula I, in an amount and under conditionssufficient to modulate metalloenzyme activity.

In one aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-related disorder ordisease, comprising administering to the subject an effective amount ofa compound or pharmaceutical composition of Formula I.

In another aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-related disorder ordisease, wherein the subject has been identified as in need of treatmentfor a metalloenzyme-related disorder or disease, comprisingadministering to said subject in need thereof, an effective amount of acompound or pharmaceutical composition of Formula I, such that saidsubject is treated for said disorder.

In another aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-mediated disorder ordisease, wherein the subject has been identified as in need of treatmentfor a metalloenzyme-mediated disorder or disease, comprisingadministering to said subject in need thereof, an effective amount of acompound or pharmaceutical composition of Formula I, such thatmetalloenzyme activity in said subject is modulated (e.g., downregulated, inhibited).

The methods herein include those wherein the disease or disorder ismediated by any of 4-hydroxyphenyl pyruvate dioxygenase, 5-lipoxygenase,adenosine deaminase, alcohol dehydrogenase, aminopeptidase N,angiotensin converting enzyme, aromatase (CYP19), calcineurin, carbamoylphosphate synthetase, carbonic anhydrase family, catechol-O-methyltransferase, cyclooxygenase family, dihydropyrimidine dehydrogenase-1,DNA polymerase, farnesyl diphosphate synthase, farnesyl transferase,fumarate reductase, GABA aminotransferase, HIF-prolyl hydroxylase,histone deacetylase family, HIV integrase, HIV-1 reverse transcriptase,isoleucine tRNA ligase, lanosterol demethylase (CYP51), matrixmetalloprotease family, methionine aminopeptidase, neutralendopeptidase, nitric oxide synthase family, phosphodiesterase III,phosphodiesterase IV, phosphodiesterase V, pyruvate ferredoxinoxidoreductase, renal peptidase, ribonucleoside diphosphate reductase,thromboxane synthase (CYP5a), thyroid peroxidase, tyrosinase, urease, orxanthine oxidase.

The methods herein include those wherein the disease or disorder ismediated by any of 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR), 17-alpha hydroxylase (CYP17), aldosterone synthase (CYP11B2),aminopeptidase P, anthrax lethal factor, arginase, beta-lactamase,cytochrome P450 2A6, D-Ala D-Ala ligase, dopamine beta-hydroxylase,endothelin converting enzyme-1, glutamate carboxypeptidase II,glutaminyl cyclase, glyoxalase, heme oxygenase, HPV/HSV E1 helicase,indoleamine 2,3-dioxygenase, leukotriene A4 hydrolase, methionineaminopeptidase 2, peptide deformylase, phosphodiesterase VII, relaxase,retinoic acid hydroxylase (CYP26), TNF-alpha converting enzyme (TACE),UDP-(3-O—(R-3-hydroxymyristoyl))-N-acetylglucosamine deacetylase (LpxC),vascular adhesion protein-1 (VAP-1), or vitamin D hydroxylase (CYP24).

The methods herein include those wherein the disease or disorder iscancer, cardiovascular disease, inflammatory disease, infectiousdisease, metabolic disease, ophthalmologic disease, central nervoussystem (CNS) disease, urologic disease, or gastrointestinal disease.

The methods herein include those wherein the disease or disorder isprostate cancer, breast cancer, inflammatory bowel disease, psoriasis,systemic fungal infection, skin structure fungal infection, mucosalfungal infection, or onychomycosis.

Methods delineated herein include those wherein the subject isidentified as in need of a particular stated treatment. Identifying asubject in need of such treatment can be in the judgment of a subject ora health care professional and can be subjective (e.g. opinion) orobjective (e.g. measurable by a test or diagnostic method).

DETAILED DESCRIPTION Definitions

In order that the invention may be more readily understood, certainterms are first defined here for convenience.

As used herein, the term “treating” a disorder encompasses preventing,ameliorating, mitigating and/or managing the disorder and/or conditionsthat may cause the disorder. The terms “treating” and “treatment” referto a method of alleviating or abating a disease and/or its attendantsymptoms. In accordance with the present invention “treating” includespreventing, blocking, inhibiting, attenuating, protecting against,modulating, reversing the effects of and reducing the occurrence ofe.g., the harmful effects of a disorder.

As used herein, “inhibiting” encompasses preventing, reducing andhalting progression. Note that “enzyme inhibition” (e.g., metalloenzymeinhibition) is distinguished and described below.

The term “modulate” refers to increases or decreases in the activity ofan enzyme in response to exposure to a compound of the invention.

The terms “isolated,” “purified,” or “biologically pure” refer tomaterial that is substantially or essentially free from components thatnormally accompany it as found in its native state. Purity andhomogeneity are typically determined using analytical chemistrytechniques such as polyacrylamide gel electrophoresis or highperformance liquid chromatography. Particularly, in embodiments thecompound is at least 85% pure, more preferably at least 90% pure, morepreferably at least 95% pure, and most preferably at least 99% pure.

The term “administration” or “administering” includes routes ofintroducing the compound(s) to a subject to perform its (their) intendedfunction. Examples of routes of administration which can be used includeinjection (subcutaneous, intravenous, parenterally, intraperitoneally,intrathecal), topical, oral, inhalation, rectal and transdermal.

The term “effective amount” includes an amount effective, at dosages andfor periods of time necessary, to achieve the desired result. Aneffective amount of compound may vary according to factors such as thedisease state, age, and weight of the subject, and the ability of thecompound to elicit a desired response in the subject. Dosage regimensmay be adjusted to provide the optimum therapeutic response. Aneffective amount is also one in which any toxic or detrimental effects(e.g., side effects) of the inhibitor compound are outweighed by thetherapeutically beneficial effects.

The phrases “systemic administration,” “administered systemically”,“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound(s), drug or other material,such that it enters the patient's system and, thus, is subject tometabolism and other like processes.

The term “therapeutically effective amount” refers to that amount of thecompound being administered sufficient to prevent development of oralleviate to some extent one or more of the symptoms of the condition ordisorder being treated.

A therapeutically effective amount of compound (i.e., an effectivedosage) may range from about 0.005 micrograms per kilogram (μg/kg) toabout 200 milligrams per kilogram (mg/kg), preferably about 0.01 mg/kgto about 200 mg/kg, more preferably about 0.015 mg/kg to about 30 mg/kgof body weight. In other embodiments, the therapeutically effect amountmay range from about 1.0 picomolar (pM) to about 10 micromolar (μM). Theskilled artisan will appreciate that certain factors may influence thedosage required to effectively treat a subject, including but notlimited to the severity of the disease or disorder, previous treatments,the general health and/or age of the subject, and other diseasespresent. Moreover, treatment of a subject with a therapeuticallyeffective amount of a compound can include a single treatment or,preferably, can include a series of treatments. In one example, asubject is treated with a compound in the range of between about 0.005μg/kg to about 200 mg/kg of body weight, one time per day for betweenabout 1 to 10 weeks, preferably between 2 to 8 weeks, more preferablybetween about 3 to 7 weeks, and even more preferably for about 4, 5, or6 weeks. In another example, a subject may be treated daily for severalyears in the setting of a chronic condition or illness. It will also beappreciated that the effective dosage of a compound used for treatmentmay increase or decrease over the course of a particular treatment.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “diastereomers” refers to stereoisomers with two or morecenters of dissymmetry and whose molecules are not mirror images of oneanother.

The term “enantiomers” refers to two stereoisomers of a compound whichare non-superimposable mirror images of one another. An equimolarmixture of two enantiomers is called a “racemic mixture” or a“racemate.”

The term “isomers” or “stereoisomers” refers to compounds which haveidentical chemical constitution, but differ with regard to thearrangement of the atoms or groups in space.

The term “prodrug” includes compounds with moieties which can bemetabolized in vivo. Generally, the prodrugs are metabolized in vivo byesterases or by other mechanisms to active drugs. Examples of prodrugsand their uses are well known in the art (See, e.g., Berge et al. (1977)“Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19). The prodrugs can beprepared in situ during the final isolation and purification of thecompounds, or by separately reacting the purified compound in its freeacid form or hydroxyl with a suitable esterifying agent. Hydroxyl groupscan be converted into esters via treatment with a carboxylic acid.Examples of prodrug moieties include substituted and unsubstituted,branched or unbranched lower alkyl ester moieties, (e.g., propionic acidesters), lower alkenyl esters, di-lower alkyl-amino lower-alkyl esters(e.g., dimethylaminoethyl ester), acylamino lower alkyl esters (e.g.,acetyloxymethyl ester), acyloxy lower alkyl esters (e.g.,pivaloyloxymethyl ester), aryl esters (phenyl ester), aryl-lower alkylesters (e.g., benzyl ester), substituted (e.g., with methyl, halo, ormethoxy substituents) aryl and aryl-lower alkyl esters, amides,lower-alkyl amides, di-lower alkyl amides, and hydroxy amides. Preferredprodrug moieties are propionic acid esters and acyl esters. Prodrugswhich are converted to active forms through other mechanisms in vivo arealso included. In aspects, the compounds of the invention are prodrugsof any of the formulae herein.

The term “subject” refers to animals such as mammals, including, but notlimited to, primates (e.g., humans), cows, sheep, goats, horses, dogs,cats, rabbits, rats, mice and the like. In certain embodiments, thesubject is a human.

The terms “a,” “an,” and “the” refer to “one or more” when used in thisapplication, including the claims. Thus, for example, reference to “asample” includes a plurality of samples, unless the context clearly isto the contrary (e.g., a plurality of samples), and so forth.

Throughout this specification and the claims, the words “comprise,”“comprises,” and “comprising” are used in a non-exclusive sense, exceptwhere the context requires otherwise.

As used herein, the term “about,” when referring to a value is meant toencompass variations of, in some embodiments ±20%, in some embodiments±10%, in some embodiments ±5%, in some embodiments ±1%, in someembodiments ±0.5%, and in some embodiments ±0.1% from the specifiedamount, as such variations are appropriate to perform the disclosedmethods or employ the disclosed compositions.

Use of the word “inhibitor” herein is meant to mean a molecule thatexhibits activity for inhibiting a metalloenzyme. By “inhibit” herein ismeant to decrease the activity of a metalloenzyme, as compared to theactivity of metalloenzyme in the absence of the inhibitor. In someembodiments, the term “inhibit” means a decrease in metalloenzymeactivity of at least about 5%, at least about 10%, at least about 20%,at least about 25%, at least about 50%, at least about 60%, at leastabout 70%, at least about 80%, at least about 90%, or at least about95%. In other embodiments, inhibit means a decrease in metalloenzymeactivity of about 5% to about 25%, about 25% to about 50%, about 50% toabout 75%, or about 75% to 100%. In some embodiments, inhibit means adecrease in metalloenzyme activity of about 95% to 100%, e.g., adecrease in activity of 95%, 96%, 97%, 98%, 99%, or 100%. Such decreasescan be measured using a variety of techniques that would be recognizableby one of skill in the art. Particular assays for measuring individualactivity are described below.

Furthermore the compounds of the invention include olefins having eithergeometry:

“Z” refers to what is referred to as a “cis” (same side) configurationwhereas “E” refers to what is referred to as a “trans” (opposite side)configuration. With respect to the nomenclature of a chiral center, theterms “d” and “1” configuration are as defined by the IUPACRecommendations. As to the use of the terms, diastereomer, racemate,epimer and enantiomer, these will be used in their normal context todescribe the stereochemistry of preparations.

As used throughout this specification, the term ‘R’ refers to the groupconsisting of C₁₋₈ alkyl, C₃₋₈ alkenyl or C₃₋₈ alkynyl, unless statedotherwise.

As used herein, the term “alkyl” refers to a straight-chained orbranched hydrocarbon group containing 1 to 12 carbon atoms. The term“lower alkyl” refers to a C₁-C₆ alkyl chain. Examples of alkyl groupsinclude methyl, ethyl, n-propyl, isopropyl, tert-butyl, and n-pentyl.Alkyl groups may be optionally substituted with one or moresubstituents.

The term “alkenyl” refers to an unsaturated hydrocarbon chain that maybe a straight chain or branched chain, containing 2 to 12 carbon atomsand at least one carbon-carbon double bond. Alkenyl groups may beoptionally substituted with one or more substituents.

The term “alkynyl” refers to an unsaturated hydrocarbon chain that maybe a straight chain or branched chain, containing 2 to 12 carbon atomsand at least one carbon-carbon triple bond. Alkynyl groups may beoptionally substituted with one or more substituents.

The sp² or sp carbons of an alkenyl group and an alkynyl group,respectively, may optionally be the point of attachment of the alkenylor alkynyl groups.

The term “alkoxy” refers to an —OR substituent where R is C₁₋₈ alkyl,C₃₋₈ alkenyl or C₃₋₈ alkynyl, unless stated otherwise.

As used herein, the term “halogen”, “hal” or “halo” means —F, —Cl, —Bror —I.

The term “haloalkoxy” refers to an —OR substituent where R is fully orpartially substituted with Cl, F, I, or Br or any combination thereof.Examples of haloalkoxy groups include trifluoromethoxy, and2,2,2-trifluoroethoxy.

The term “cycloalkyl” refers to a hydrocarbon 3-8 membered monocyclic or7-14 membered bicyclic ring system having at least one saturated ring orhaving at least one non-aromatic ring, wherein the non-aromatic ring mayhave some degree of unsaturation. Cycloalkyl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, or 4 atoms of each ring of a cycloalkyl group may be substituted by asubstituent. Representative examples of a cycloalkyl group includecyclopropyl, cyclopentyl, cyclohexyl, cyclobutyl, cycloheptyl,cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and thelike.

The term “aryl” refers to a hydrocarbon monocyclic, bicyclic ortricyclic aromatic ring system. Aryl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, 4, 5 or 6 atoms of each ring of an aryl group may be substituted by asubstituent. Examples of aryl groups include phenyl, naphthyl,anthracenyl, fluorenyl, indenyl, azulenyl, and the like.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic,8-12 membered bicyclic, or 11-14 membered tricyclic ring system having1-4 ring heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms selected from O, N, or S, andthe remainder ring atoms being carbon (with appropriate hydrogen atomsunless otherwise indicated). Heteroaryl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, or 4 atoms of each ring of a heteroaryl group may be substituted by asubstituent. Examples of heteroaryl groups include pyridyl, furanyl,thienyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, thiazolyl,isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridazinyl,pyrimidinyl, pyrazinyl, triazinyl, isoquinolinyl, indazolyl, and thelike.

The term “nitrogen-containing heteroaryl” refers to a heteroaryl grouphaving 1-4 ring nitrogen heteroatoms if monocyclic, 1-6 ring nitrogenheteroatoms if bicyclic, or 1-9 ring nitrogen heteroatoms if tricyclic.

The term “heterocycloalkyl” refers to a nonaromatic 3-8 memberedmonocyclic, 7-12 membered bicyclic, or 10-14 membered tricyclic ringsystem comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms ifbicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selectedfrom O, N, S, B, P or Si, wherein the nonaromatic ring system iscompletely saturated. Heterocycloalkyl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, or 4 atoms of each ring of a heterocycloalkyl group may besubstituted by a substituent. Representative heterocycloalkyl groupsinclude piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl,thiomorpholinyl, 1,3-dioxolane, tetrahydrofuranyl, tetrahydrothienyl,thiirenyl, and the like.

The term “alkylamino” refers to an amino substituent which is furthersubstituted with one or two alkyl groups. The term “aminoalkyl” refersto an alkyl substituent which is further substituted with one or moreamino groups. The term “hydroxyalkyl” or “hydroxylalkyl” refers to analkyl substituent which is further substituted with one or more hydroxylgroups. The alkyl or aryl portion of alkylamino, aminoalkyl,mercaptoalkyl, hydroxyalkyl, mercaptoalkoxy, sulfonylalkyl,sulfonylaryl, alkylcarbonyl, and alkylcarbonylalkyl may be optionallysubstituted with one or more substituents.

Acids and bases useful in the methods herein are known in the art. Acidcatalysts are any acidic chemical, which can be inorganic (e.g.,hydrochloric, sulfuric, nitric acids, aluminum trichloride) or organic(e.g., camphorsulfonic acid, p-toluenesulfonic acid, acetic acid,ytterbium triflate) in nature. Acids are useful in either catalytic orstoichiometric amounts to facilitate chemical reactions. Bases are anybasic chemical, which can be inorganic (e.g., sodium bicarbonate,potassium hydroxide) or organic (e.g., triethylamine, pyridine) innature. Bases are useful in either catalytic or stoichiometric amountsto facilitate chemical reactions.

Alkylating agents are any reagent that is capable of effecting thealkylation of the functional group at issue (e.g., oxygen atom of analcohol, nitrogen atom of an amino group). Alkylating agents are knownin the art, including in the references cited herein, and include alkylhalides (e.g., methyl iodide, benzyl bromide or chloride), alkylsulfates (e.g., methyl sulfate), or other alkyl group-leaving groupcombinations known in the art. Leaving groups are any stable speciesthat can detach from a molecule during a reaction (e.g., eliminationreaction, substitution reaction) and are known in the art, including inthe references cited herein, and include halides (e.g., I—, Cl—, Br—,F—), hydroxy, alkoxy (e.g., —OMe, —O-t-Bu), acyloxy anions (e.g., —OAc,—OC(O)CF₃), sulfonates (e.g., mesyl, tosyl), acetamides (e.g.,—NHC(O)Me), carbamates (e.g., N(Me)C(O)Ot-Bu), phosphonates (e.g.,—OP(O)(OEt)₂), water or alcohols (protic conditions), and the like.

In certain embodiments, substituents on any group (such as, for example,alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl,cycloalkyl, heterocycloalkyl) can be at any atom of that group, whereinany group that can be substituted (such as, for example, alkyl, alkenyl,alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl,heterocycloalkyl) can be optionally substituted with one or moresubstituents (which may be the same or different), each replacing ahydrogen atom. Examples of suitable substituents include, but are notlimited to alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano,nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl),carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl,alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl,thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl,dialkylamino, alkylcarbonylamino, alkylaminocarbonyl,alkoxycarbonylamino, alkylamino, arylamino, diarylamino, alkylcarbonyl,or arylamino-substituted aryl; arylalkylamino, aralkylaminocarbonyl,amido, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl,alkylsulfonylamino, arylsulfonylamino, imino, carbamido, carbamyl,thioureido, thiocyanato, sulfoamido, sulfonylalkyl, sulfonylaryl,mercaptoalkoxy, N-hydroxyamidinyl, or N′-aryl, N″-hydroxyamidinyl.

Compounds of the invention can be made by means known in the art oforganic synthesis. Methods for optimizing reaction conditions, ifnecessary minimizing competing by-products, are known in the art.Reaction optimization and scale-up may advantageously utilize high-speedparallel synthesis equipment and computer-controlled microreactors (e.g.Design And Optimization in Organic Synthesis, 2^(nd) Edition, Carlson R,Ed, 2005; Elsevier Science Ltd.; Jähnisch, K. et al., Angew. Chem. Int.Ed. Engl. 2004, 43, 406; and references therein). Additional reactionschemes and protocols may be determined by the skilled artisan by use ofcommercially available structure-searchable database software, forinstance, SciFinder® (CAS division of the American Chemical Society) andCrossFire Beilstein® (Elsevier MDL), or by appropriate keyword searchingusing an internet search engine such as Google® or keyword databasessuch as the US Patent and Trademark Office text database.

The compounds herein may also contain linkages (e.g., carbon-carbonbonds) wherein bond rotation is restricted about that particularlinkage, e.g. restriction resulting from the presence of a ring ordouble bond. Accordingly, all cis/trans and E/Z isomers are expresslyincluded in the present invention. The compounds herein may also berepresented in multiple tautomeric forms; in such instances, theinvention expressly includes all tautomeric forms of the compoundsdescribed herein, even though only a single tautomeric form may berepresented. All such isomeric forms of such compounds herein areexpressly included in the present invention. All crystal forms andpolymorphs of the compounds described herein are expressly included inthe present invention. Also embodied are extracts and fractionscomprising compounds of the invention. The term isomers is intended toinclude diastereoisomers, enantiomers, regioisomers, structural isomers,rotational isomers, tautomers, and the like. For compounds which containone or more stereogenic centers, e.g., chiral compounds, the methods ofthe invention may be carried out with an enantiomerically enrichedcompound, a racemate, or a mixture of diastereomers.

Preferred enantiomerically enriched compounds have an enantiomericexcess of 50% or more, more preferably the compound has an enantiomericexcess of 60%, 70%, 80%, 90%, 95%, 98%, or 99% or more. In preferredembodiments, only one enantiomer or diastereomer of a chiral compound ofthe invention is administered to cells or a subject.

Methods of Treatment

In one aspect, the invention provides a method of modulating themetalloenzyme activity of a cell in a subject, comprising contacting thesubject with a compound of Formula I, in an amount and under conditionssufficient to modulate metalloenzyme activity.

In one embodiment, the modulation is inhibition.

In another aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-mediated disorder ordisease, comprising administering to the subject an effective amount ofa compound or pharmaceutical composition of Formula I.

In other aspects, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-mediated disorder ordisease, wherein the subject has been identified as in need of treatmentfor a metalloenzyme-mediated disorder or disease, comprisingadministering to said subject in need thereof, an effective amount of acompound or pharmaceutical composition of Formula I, such that saidsubject is treated for said disorder.

In certain embodiments, the invention provides a method of treating adisease, disorder or symptom thereof, wherein the disorder is cancer,cardiovascular disease, inflammatory disease or infectious disease. Inother embodiments the disease, disorder or symptom thereof is metabolicdisease, ophthalmologic disease, central nervous system (CNS) disease,urologic disease, or gastrointestinal disease. In certain embodimentsthe disease is prostate cancer, breast cancer, inflammatory boweldisease, psoriasis, systemic fungal infection, skin structure fungalinfection, mucosal fungal infection, and onychomycosis.

In another aspect, the compounds and compositions herein are useful fortreating a disease, disorder or symptom thereof, which is associatedwith one or more of the following pathogenic fungi: Absidia corymbifera,Ajellornyces dermatitides, Arthroderma benhamiae, Arthroderma fulvum,Arthroderma gypseum, Arthroderma incurvaturn, Arthroderma otae,Arthroderma vanbreuseghemii, Aspergillus flavus, Aspergillus fumigates,Aspergillus niger, Blastomyces dermatitides, Candida albicans, Candidaglabrata, Candida guilliermondii, Candida krusei, Candida parapsilosis,Candida tropicalis, Candida pelliculosa, Cladophialophora carrionii,Coccidioides immitis, Cryptococcus neoformans, Cunninghamella sp.,Epidermophyton floccosum, Exophiala dermatitides, Filobasidiellaneoformans, Fonsecaea pedrosoi, Fusarium solani, Geotrichum candidum,Histoplasma capsulaturn, Hortaea werneckii, Issatschenkia orientalis,Madurella grisae, Malassezia fur fur, Malassezia globosa, Malasseziaobtusa, Malassezia pachydermatis, Malassezia restricta, Malasseziaslooffiae, Malassezia sympodialis, Microsporum canis, Microsporumfulvum, Microsporum gypseum, Mucor circinelloides, Nectria haematococca,Paecilomyces variotii, Paracoccidioides brasiliensis, Penicilliummarneffei, Pichia anomala, Pichia guilliermondii, Pneumocystis carinii,Pseudallescheria boydii, Rhizopus oryzae, Rhodotorula rubra,Scedosporium apiospernium, Schizophyllum commune, Sporothrix schenckii,Trichophyton mentagrophytes, Trichophyton rubrum, Trichophytonverrucosum, Trichophyton violaceum, Trichosporon asahii, Trichosporoncutaneum, Trichosporon inkin, Trichosporon mucoides.

In another aspect, the compounds and compositions herein are useful fortreating a disease, disorder or symptom thereof, which is associatedwith one of the following conditions: Aspergillosis, Blastomycosis,Candidiasis, Chromomycosis, Coccidioidomycosis, Cryptococcosis,Dermatophytoses, Histoplasmosis, Keratomycosis, Lobomycosis, Malasseziainfection, Mucormycosis, Paracoccidioidomycosis, Penicillium marneffeiinfection, Phaeohyphomycosis, Pneumocystis pneumonia, Rhinosporidiosis.

In another aspect, the compounds and compositions herein are useful fortreating a disease, disorder or symptom thereof, which is Chagas disease(Genus Trypanosoma), African trypanosomiasis (Genus Trypanosoma),leishmaniasis (Genus Leishmania), tuberculosis (Genus Mycobacterium),leprosy (Genus Mycobacterium), malaria (Genus Plasmodium), tinea(capitis, corporis, pedis, tonsurans, versicolor).

In certain embodiments, the subject is a mammal, preferably a primate orhuman.

In another embodiment, the invention provides a method as describedabove, wherein the effective amount of the compound of Formula I is asdescribed above.

In another embodiment, the invention provides a method as describedabove, wherein the compound of Formula I is administered intravenously,intramuscularly, subcutaneously, intracerebroventricularly, orally ortopically.

In another embodiment, the invention provides a method as describedherein wherein the compound of Formula I demonstrates selectivity for anactivity range against a target organism or enzyme and an activity rangeagainst an off-target enzyme (e.g., C. albicans MIC<0.02 μg/mL andIC₅₀>16 μM for CYP2C₉, CYP2C19 and CYP3A4; C. albicans MIC<0.10 μg/mLand IC₅₀>10 μM for CYP2C9, CYP2C19 and CYP3A4; C. albicans MIC<0.5 μg/mLand IC₅₀>15 μM for CYP2C9, CYP2C19 and CYP3A4).

In other embodiments, the invention provides a method as describedabove, wherein the compound of Formula I is administered alone or incombination with one or more other therapeutics. In a furtherembodiment, the additional therapeutic agent is an anti-cancer agent,antifungal agent, cardiovascular agent, anti-inflammatory agent,chemotherapeutic agent, an anti-angiogenesis agent, cytotoxic agent, ananti-proliferation agent, metabolic disease agent, ophthalmologicdisease agent, central nervous system (CNS) disease agent, urologicdisease agent, or gastrointestinal disease agent.

Another object of the present invention is the use of a compound asdescribed herein (e.g., of any formulae herein) in the manufacture of amedicament for use in the treatment of a metalloenzyme-mediated disorderor disease. Another object of the present invention is the use of acompound as described herein (e.g., of any formulae herein) for use inthe treatment of a metalloenzyme-mediated disorder or disease.

Pharmaceutical Compositions

In one aspect, the invention provides a pharmaceutical compositioncomprising the compound of Formula I and a pharmaceutically acceptablecarrier.

In another embodiment, the invention provides a pharmaceuticalcomposition further comprising an additional therapeutic agent. In afurther embodiment, the additional therapeutic agent is an anti-canceragent, antifungal agent, cardiovascular agent, anti-inflammatory agent,chemotherapeutic agent, an anti-angiogenesis agent, cytotoxic agent, ananti-proliferation agent, metabolic disease agent, ophthalmologicdisease agent, central nervous system (CNS) disease agent, urologicdisease agent, or gastrointestinal disease agent.

In one aspect, the invention provides a kit comprising an effectiveamount of a compound of Formula I, in unit dosage form, together withinstructions for administering the compound to a subject suffering fromor susceptible to a metalloenzyme-mediated disease or disorder,including cancer, solid tumor, cardiovascular disease, inflammatorydisease, infectious disease. In other embodiments the disease, disorderor symptom thereof is metabolic disease, ophthalmologic disease, centralnervous system (CNS) disease, urologic disease, or gastrointestinaldisease.

The term “pharmaceutically acceptable salts” or “pharmaceuticallyacceptable carrier” is meant to include salts of the active compoundswhich are prepared with relatively nontoxic acids or bases, depending onthe particular substituents found on the compounds described herein.When compounds of the present invention contain relatively acidicfunctionalities, base addition salts can be obtained by contacting theneutral form of such compounds with a sufficient amount of the desiredbase, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable base addition salts include sodium,potassium, calcium, ammonium, organic amino, or magnesium salt, or asimilar salt. When compounds of the present invention contain relativelybasic functionalities, acid addition salts can be obtained by contactingthe neutral form of such compounds with a sufficient amount of thedesired acid, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable acid addition salts include those derivedfrom inorganic acids like hydrochloric, hydrobromic, nitric, carbonic,monohydrogencarbonic, phosphoric, monohydrogenphosphoric,dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydroiodic, orphosphorous acids and the like, as well as the salts derived fromrelatively nontoxic organic acids like acetic, propionic, isobutyric,maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic,phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric,methanesulfonic, and the like. Also included are salts of amino acidssuch as arginate and the like, and salts of organic acids likeglucuronic or galactunoric acids and the like (see, e.g., Berge et al.,Journal of Pharmaceutical Science 66:1-19 (1977)). Certain specificcompounds of the present invention contain both basic and acidicfunctionalities that allow the compounds to be converted into eitherbase or acid addition salts. Other pharmaceutically acceptable carriersknown to those of skill in the art are suitable for the presentinvention.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

The invention also provides a pharmaceutical composition, comprising aneffective amount a compound described herein and a pharmaceuticallyacceptable carrier. In an embodiment, compound is administered to thesubject using a pharmaceutically-acceptable formulation, e.g., apharmaceutically-acceptable formulation that provides sustained deliveryof the compound to a subject for at least 12 hours, 24 hours, 36 hours,48 hours, one week, two weeks, three weeks, or four weeks after thepharmaceutically-acceptable formulation is administered to the subject.

Actual dosage levels and time course of administration of the activeingredients in the pharmaceutical compositions of this invention may bevaried so as to obtain an amount of the active ingredient which iseffective to achieve the desired therapeutic response for a particularpatient, composition, and mode of administration, without being toxic(or unacceptably toxic) to the patient.

In use, at least one compound according to the present invention isadministered in a pharmaceutically effective amount to a subject in needthereof in a pharmaceutical carrier by intravenous, intramuscular,subcutaneous, or intracerebroventricular injection or by oraladministration or topical application. In accordance with the presentinvention, a compound of the invention may be administered alone or inconjunction with a second, different therapeutic.

By “in conjunction with” is meant together, substantially simultaneouslyor sequentially. In one embodiment, a compound of the invention isadministered acutely. The compound of the invention may therefore beadministered for a short course of treatment, such as for about 1 day toabout 1 week. In another embodiment, the compound of the invention maybe administered over a longer period of time to ameliorate chronicdisorders, such as, for example, for about one week to several monthsdepending upon the condition to be treated.

By “pharmaceutically effective amount” as used herein is meant an amountof a compound of the invention, high enough to significantly positivelymodify the condition to be treated but low enough to avoid serious sideeffects (at a reasonable benefit/risk ratio), within the scope of soundmedical judgment. A pharmaceutically effective amount of a compound ofthe invention will vary with the particular goal to be achieved, the ageand physical condition of the patient being treated, the severity of theunderlying disease, the duration of treatment, the nature of concurrenttherapy and the specific compound employed. For example, atherapeutically effective amount of a compound of the inventionadministered to a child or a neonate will be reduced proportionately inaccordance with sound medical judgment. The effective amount of acompound of the invention will thus be the minimum amount which willprovide the desired effect.

A decided practical advantage of the present invention is that thecompound may be administered in a convenient manner such as byintravenous, intramuscular, subcutaneous, oral orintracerebroventricular injection routes or by topical application, suchas in creams or gels. Depending on the route of administration, theactive ingredients which comprise a compound of the invention may berequired to be coated in a material to protect the compound from theaction of enzymes, acids and other natural conditions which mayinactivate the compound. In order to administer a compound of theinvention by other than parenteral administration, the compound can becoated by, or administered with, a material to prevent inactivation.

The compound may be administered parenterally or intraperitoneally.Dispersions can also be prepared, for example, in glycerol, liquidpolyethylene glycols, and mixtures thereof, and in oils.

Some examples of substances which can serve as pharmaceutical carriersare sugars, such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethylcellulose, ethylcellulose and cellulose acetates; powderedtragancanth; malt; gelatin; talc; stearic acids; magnesium stearate;calcium sulfate; vegetable oils, such as peanut oils, cottonseed oil,sesame oil, olive oil, corn oil and oil of theobroma; polyols such aspropylene glycol, glycerine, sorbitol, mannitol, and polyethyleneglycol; agar; alginic acids; pyrogen-free water; isotonic saline; andphosphate buffer solution; skim milk powder; as well as other non-toxiccompatible substances used in pharmaceutical formulations such asVitamin C, estrogen and Echinacea, for example.

Wetting agents and lubricants such as sodium lauryl sulfate, as well ascoloring agents, flavoring agents, lubricants, excipients, tabletingagents, stabilizers, antioxidants and preservatives, can also bepresent. Solubilizing agents, including for example, cremaphore andbeta-cyclodextrins can also used in the pharmaceutical compositionsherein.

Pharmaceutical compositions comprising the active compounds of thepresently disclosed subject matter (or prodrugs thereof) can bemanufactured by means of conventional mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping orlyophilization processes. The compositions can be formulated inconventional manner using one or more physiologically acceptablecarriers, diluents, excipients or auxiliaries which facilitateprocessing of the active compounds into preparations which can be usedpharmaceutically.

Pharmaceutical compositions of the presently disclosed subject mattercan take a form suitable for virtually any mode of administration,including, for example, topical, ocular, oral, buccal, systemic, nasal,injection, transdermal, rectal, vaginal, and the like, or a formsuitable for administration by inhalation or insufflation.

For topical administration, the active compound(s) or prodrug(s) can beformulated as solutions, gels, ointments, creams, suspensions, and thelike.

Systemic formulations include those designed for administration byinjection, e.g., subcutaneous, intravenous, intramuscular, intrathecalor intraperitoneal injection, as well as those designed for transdermal,transmucosal, oral, or pulmonary administration.

Useful injectable preparations include sterile suspensions, solutions oremulsions of the active compound(s) in aqueous or oily vehicles. Thecompositions also can contain formulating agents, such as suspending,stabilizing and/or dispersing agent. The formulations for injection canbe presented in unit dosage form (e.g., in ampules or in multidosecontainers) and can contain added preservatives.

Alternatively, the injectable formulation can be provided in powder formfor reconstitution with a suitable vehicle, including but not limited tosterile pyrogen-free water, buffer, dextrose solution, and the like,before use. To this end, the active compound(s) can be dried by anyart-known technique, such as lyophilization, and reconstituted prior touse.

For transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants are knownin the art.

For oral administration, the pharmaceutical compositions can take theform of, for example, lozenges, tablets or capsules prepared byconventional means with pharmaceutically acceptable excipients such asbinding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidoneor hydroxypropyl methylcellulose); fillers (e.g., lactose,microcrystalline cellulose or calcium hydrogen phosphate); lubricants(e.g., magnesium stearate, talc or silica); disintegrants (e.g., potatostarch or sodium starch glycolate); or wetting agents (e.g., sodiumlauryl sulfate). The tablets can be coated by methods well known in theart with, for example, sugars or enteric coatings.

Liquid preparations for oral administration can take the form of, forexample, elixirs, solutions, syrups or suspensions, or they can bepresented as a dry product for constitution with water or other suitablevehicle before use. Such liquid preparations can be prepared byconventional means with pharmaceutically acceptable additives such assuspending agents (e.g., sorbitol syrup, cellulose derivatives orhydrogenated edible fats); emulsifying agents (e.g., lecithin oracacia); non aqueous vehicles (e.g., almond oil, oily esters, ethylalcohol or fractionated vegetable oils); and preservatives (e.g., methylor propylp-hydroxybenzoates or sorbic acid). The preparations also cancontain buffer salts, preservatives, flavoring, coloring and sweeteningagents as appropriate.

Preparations for oral administration can be suitably formulated to givecontrolled release of the active compound or prodrug, as is well known.

For buccal administration, the compositions can take the form of tabletsor lozenges formulated in a conventional manner.

For rectal and vaginal routes of administration, the active compound(s)can be formulated as solutions (for retention enemas), suppositories, orointments containing conventional suppository bases, such as cocoabutter or other glycerides.

For nasal administration or administration by inhalation orinsufflation, the active compound(s) or prodrug(s) can be convenientlydelivered in the form of an aerosol spray from pressurized packs or anebulizer with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or othersuitable gas. In the case of a pressurized aerosol, the dosage unit canbe determined by providing a valve to deliver a metered amount. Capsulesand cartridges for use in an inhaler or insufflator (for examplecapsules and cartridges comprised of gelatin) can be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

A specific example of an aqueous suspension formulation suitable fornasal administration using commercially-available nasal spray devicesincludes the following ingredients: active compound or prodrug (0.5-20mg/mL); benzalkonium chloride (0.1-0.2 mg/mL); polysorbate 80 (TWEEN®80; 0.5-5 mg/mL); carboxymethylcellulose sodium or microcrystallinecellulose (1-15 mg/mL); phenylethanol (1-4 mg/mL); and dextrose (20-50mg/mL). The pH of the final suspension can be adjusted to range fromabout pH 5 to pH 7, with a pH of about pH 5.5 being typical.

For ocular administration, the active compound(s) or prodrug(s) can beformulated as a solution, emulsion, suspension, and the like, suitablefor administration to the eye. A variety of vehicles suitable foradministering compounds to the eye are known in the art. Specificnon-limiting examples are described in U.S. Pat. Nos. 6,261,547;6,197,934; 6,056,950; 5,800,807; 5,776,445; 5,698,219; 5,521,222;5,403,841; 5,077,033; 4,882,150; and 4,738,851, each of which isincorporated herein by reference in its entirety.

For prolonged delivery, the active compound(s) or prodrug(s) can beformulated as a depot preparation for administration by implantation orintramuscular injection. The active ingredient can be formulated withsuitable polymeric or hydrophobic materials (e.g., as an emulsion in anacceptable oil) or ion exchange resins, or as sparingly solublederivatives, e.g., as a sparingly soluble salt. Alternatively,transdermal delivery systems manufactured as an adhesive disc or patchwhich slowly releases the active compound(s) for percutaneous absorptioncan be used. To this end, permeation enhancers can be used to facilitatetransdermal penetration of the active compound(s). Suitable transdermalpatches are described in for example, U.S. Pat. Nos. 5,407,713;5,352,456; 5,332,213; 5,336,168; 5,290,561; 5,254,346; 5,164,189;5,163,899; 5,088,977; 5,087,240; 5,008,110; and 4,921,475, each of whichis incorporated herein by reference in its entirety.

Alternatively, other pharmaceutical delivery systems can be employed.Liposomes and emulsions are well-known examples of delivery vehiclesthat can be used to deliver active compound(s) or prodrug(s). Certainorganic solvents such as dimethylsulfoxide (DMSO) also can be employed.

The pharmaceutical compositions can, if desired, be presented in a packor dispenser device which can contain one or more unit dosage formscontaining the active compound(s).

The pack can, for example, comprise metal or plastic foil, such as ablister pack. The pack or dispenser device can be accompanied byinstructions for administration.

The active compound(s) or prodrug(s) of the presently disclosed subjectmatter, or compositions thereof, will generally be used in an amounteffective to achieve the intended result, for example in an amounteffective to treat or prevent the particular disease being treated. Thecompound(s) can be administered therapeutically to achieve therapeuticbenefit or prophylactically to achieve prophylactic benefit. Bytherapeutic benefit is meant eradication or amelioration of theunderlying disorder being treated and/or eradication or amelioration ofone or more of the symptoms associated with the underlying disorder suchthat the patient reports an improvement in feeling or condition,notwithstanding that the patient can still be afflicted with theunderlying disorder. For example, administration of a compound to apatient suffering from an allergy provides therapeutic benefit not onlywhen the underlying allergic response is eradicated or ameliorated, butalso when the patient reports a decrease in the severity or duration ofthe symptoms associated with the allergy following exposure to theallergen. As another example, therapeutic benefit in the context ofasthma includes an improvement in respiration following the onset of anasthmatic attack, or a reduction in the frequency or severity ofasthmatic episodes. Therapeutic benefit also includes halting or slowingthe progression of the disease, regardless of whether improvement isrealized.

For prophylactic administration, the compound can be administered to apatient at risk of developing one of the previously described diseases.A patient at risk of developing a disease can be a patient havingcharacteristics placing the patient in a designated group of at riskpatients, as defined by an appropriate medical professional or group. Apatient at risk may also be a patient that is commonly or routinely in asetting where development of the underlying disease that may be treatedby administration of a metalloenzyme inhibitor according to theinvention could occur. In other words, the at risk patient is one who iscommonly or routinely exposed to the disease or illness causingconditions or may be acutely exposed for a limited time. Alternatively,prophylactic administration can be applied to avoid the onset ofsymptoms in a patient diagnosed with the underlying disorder.

The amount of compound administered will depend upon a variety offactors, including, for example, the particular indication beingtreated, the mode of administration, whether the desired benefit isprophylactic or therapeutic, the severity of the indication beingtreated and the age and weight of the patient, the bioavailability ofthe particular active compound, and the like. Determination of aneffective dosage is well within the capabilities of those skilled in theart.

Effective dosages can be estimated initially from in vitro assays. Forexample, an initial dosage for use in animals can be formulated toachieve a circulating blood or serum concentration of active compoundthat is at or above an IC₅₀ of the particular compound as measured in anin vitro assay, such as the in vitro fungal MIC or MFC and other invitro assays described in the Examples section. Calculating dosages toachieve such circulating blood or serum concentrations taking intoaccount the bioavailability of the particular compound is well withinthe capabilities of skilled artisans. For guidance, see Fingl &Woodbury, “General Principles,” In: Goodman and Gilman's ThePharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46, latestedition, Pagamonon Press, and the references cited therein, which areincorporated herein by reference.

Initial dosages also can be estimated from in vivo data, such as animalmodels. Animal models useful for testing the efficacy of compounds totreat or prevent the various diseases described above are well-known inthe art.

Dosage amounts will typically be in the range of from about 0.0001 or0.001 or 0.01 milligrams per kilogram per day (mg/kg/day) to about 100mg/kg/day, but can be higher or lower, depending upon, among otherfactors, the activity of the compound, its bioavailability, the mode ofadministration, and various factors discussed above. Dosage amount andinterval can be adjusted individually to provide plasma levels of thecompound(s) which are sufficient to maintain therapeutic or prophylacticeffect. In cases of local administration or selective uptake, such aslocal topical administration, the effective local concentration ofactive compound(s) cannot be related to plasma concentration. Skilledartisans will be able to optimize effective local dosages without undueexperimentation.

The compound(s) can be administered once per day, a few or several timesper day, or even multiple times per day, depending upon, among otherthings, the indication being treated and the judgment of the prescribingphysician.

Preferably, the compound(s) will provide therapeutic or prophylacticbenefit without causing substantial toxicity. Toxicity of thecompound(s) can be determined using standard pharmaceutical procedures.The dose ratio between toxic and therapeutic (or prophylactic) effect isthe therapeutic index. Compounds(s) that exhibit high therapeuticindices are preferred.

The recitation of a listing of chemical groups in any definition of avariable herein includes definitions of that variable as any singlegroup or combination of listed groups. The recitation of an embodimentfor a variable herein includes that embodiment as any single embodimentor in combination with any other embodiments or portions thereof. Therecitation of an embodiment herein includes that embodiment as anysingle embodiment or in combination with any other embodiments orportions thereof.

EXAMPLES

The present invention will now be demonstrated using specific examplesthat are not to be construed as limiting.

General Experimental Procedures

Definitions of variables in the structures in schemes herein arecommensurate with those of corresponding positions in the formulaedelineated herein.

Synthesis of Antifungals

Syntheses of azole targets (I) may be accomplished using the examplesynthesis that is shown below (Scheme 1). A broad range of arenes andheterocycles, in addition to the 2-pyridine example below, may beprepared starting from functionalized halo-aromatic starting materials(e.g. 1). For the purpose of this example, R4 is a halogenated benzenemoiety. An example synthesis of targets (I) commences with condensationof A with copper-activated ethyl α-bromo-difluoroacetate followed bycondensation of the incipient ethyl ester product with lithiatedbromodifluorobenzene to furnish ketone B (Scheme 1). The ketone isepoxidized with diazomethane to afford C. The bromo-pyridineintermediate C may be treated with aryl-boronic acids to introduce theR3-Ph moiety of D. The product D is obtained by then opening the epoxidewith azole in the presence of a base such as potassium carbonate.

Synthesis of2-(5-Bromopyridin-2-yl)-1-(2,4-difluorophenyl)-2,2-difluoroethanone (B)

To a suspension of copper powder (2.68 grams (g), 42.2 millimoles(mmol)) in dimethyl sulfoxide (DMSO; 35 milliliters (mL)) was addedethyl bromodifluoroacetate (2.70 mL, 21.10 mmol), and the mixture wasstirred for 1 hour (h) at room temperature (RT). 2,5-Dibromopyridine(2.50 g, 10.55 mmol) was then added and stirring was continued for 15 hat RT. The reaction was quenched with aqueous ammonium chloride (NH₄Cl)and was extracted with dichloromethane (CH₂Cl₂; 3×25 mL). The combinedorganic layers were washed with water, washed with brine, dried overanhydrous sodium sulfate (Na₂SO₄), and concentrated under reducedpressure to afford crude product mixture which upon column purificationusing ethyl acetate (EtOAc)/hexane afforded the ethyl ester intermediate(2.40 g, 8.57 mmol, 81%) as a pale yellow oil. ¹H NMR (500 MHz, CDCl₃):δ 8.71 (s, 1H), 8.00 (d, J=9.0 Hz, 1H), 7.64 (d, J=9.0 Hz, 1H),4.42-4.35 (m, 2H), 1.39-1.31 (m, 3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (1.65 g, 8.57 mmol)in diethyl ether (10 mL) was added n-butyllithium (n-BuLi; 3.70 mL, 8.57mmol) at −70° C. followed by addition of the ethyl ester from above(2.40 g, 8.57 mmol) in diethyl ether (5 mL) after 15 minutes (min). Thereaction mixture was stirred for 1 h at −70° C. and was warmed to roomtemperature at which point the mixture was stirred for another 2 h. Thereaction was quenched with aqueous NH₄Cl solution and extracted withEtOAc (3×20 mL). The combined organic layers were washed with water,washed with brine, dried over anhydrous Na₂SO₄, and concentrated underreduced pressure. The crude compound was purified by columnchromatography to afford ketone B (1.30 g, 3.73 mmol, 43%) as a yellowliquid. ¹H NMR (500 MHz, CDCl₃): δ 8.62 (s, 1H), 8.08-8.04 (m, 2H),7.74-7.70 (m, 1H), 7.05-6.95 (m, 1H), 6.88-6.78 (m, 1H). MS (ESI): m/z347, 349 [(M⁺+1)+2].

5-Bromo-2-((2-(2,4-difluorophenyl)oxiran-2-yl)difluoromethyl)pyridine(C)

To a stirred solution of ketone B (1.30 g, 3.73 mmol) in diethyl ether(300 mL) was added freshly prepared diazomethane at 0° C. followed bywarming to RT. The reaction mixture was stirred for 2 h. The volatileswere removed under reduced pressure to afford a crude product mixturewhich upon column chromatography using EtOAc/hexane as the eluentafforded oxirane C (800 mg, 2.20 mmol, 59%) as light yellow solid. ¹HNMR (500 MHz, CDCl₃): δ 8.72 (s, 1H), 7.89 (d, J=9.0 Hz, 1H), 7.39-7.35(m, 2H), 6.86-6.83 (m, 1H), 6.77-6.74 (m, 1H), 3.44 (s, 1H), 2.98 (s,1H). MS (ESI): m/z 362, 364 [(M⁺+1)+2].

Example 1

4-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)benzonitrile(1)

To a stirred solution of epoxide C (0.3 g, 0.82 mmol) and4-cyano-benzene boronic acid (0.14 g, 0.99 mmol) in 1,4-dioxane (5 mL)was added potassium carbonate (K₂CO₃; 0.17 g, 1.24 mmol) at RT under aninert atmosphere. After purging with argon for a period of 30 min,1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)(Pd(dppf)₂Cl₂; 30 mg, 0.041 mmol) was added to the reaction mixtureunder argon atmosphere. The resulting mixture was stirred for 8 h at 75°C. Progress of the reaction was monitored by thin layer chromatography(TLC). The solvent was evaporated under reduced pressure; the obtainedresidue was dissolved in water (20 mL). The aqueous layer was extractedwith EtOAc (3×50 mL). The combined organic phases were washed with waterand brine, dried over anhydrous Na₂SO₄ and concentrated. The crudematerial was purified by column chromatography to afford coupled product(0.15 g, 0.39 mmol, 47%) as a solid. ¹H NMR (500 MHz, CDCl₃): δ 8.87 (s,1H), 7.95 (dd, J=8.0, 2.0 Hz, 1H), 7.81-7.77 (m, 2H), 7.71-7.68 (m, 2H),7.61 (d, J=8.0 Hz, 1H), 7.43 (app q, 1H), 6.87-6.83 (m, 1H), 6.77-6.73(m, 1H), 3.48 (d, J=5.0 Hz, 1H), 3.00 (app s, 1H). MS (ESI): m/z 385[M⁺+1].

To a stirred solution of the coupled product (150 mg, 0.39 mmol) inN,N-dimethylformamide (DMF; 3 mL) were added 1H-tetrazole (33 mg, 0.46mmol) followed by K₂CO₃ (27 mg, 0.19 mmol) at RT under an inertatmosphere. The reaction mixture was stirred for 16 h at 70° C. Thereaction mixture was cooled to RT, diluted with water (5 mL) andextracted with EtOAc (2×20 mL). The organic layer was washed with waterand brine and dried over anhydrous Na₂SO₄. After filtering off thesolid, the solvent was evaporated under reduced pressure to give crudecompound. The crude compound was purified by column chromatography toafford compound 1 (50 mg, 0.11 mmol, 28%) as a white solid. ¹H NMR (500MHz, CDCl₃): δ 8.75 (s, 1H), 8.71 (s, 1H), 8.00 (dd, J=8.0, 2.0 Hz, 1H),7.82 (d, J=7.0 Hz, 2H), 7.72 (d, J=8.5 Hz, 1H), 7.67 (d, J=7.0 Hz, 2H),7.44-7.39 (m, 1H), 7.37 (s, 1H), 6.81-6.77 (m, 1H), 6.72-6.68 (m, 1H),5.53 (d, J=14.5 Hz, 1H), 5.20 (d, J=14.5 Hz, 1H). HPLC: 99.6%. MS (ESI):m/z 455 [M⁺+1].

Example 2

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethyl)phenyl)pyridin-2-yl)propan-2-ol (2)

To a stirred solution of bromo-epoxide C (0.25 g, 0.69 mmol) intetrahydrofuran (THF; 20 mL) and water (7 mL) were added4-(trifluoromethyl)phenylboronic acid (0.10 g, 0.55 mmol), sodiumcarbonate (Na₂CO₃; 0.16 g, 1.55 mmol) and Pd(dppf)₂Cl₂ (0.14 g, 0.17mmol) at RT under an inert atmosphere. After purging with argon for aperiod of 30 min, the reaction mixture was heated to 75° C. and stirringwas continued for 4 h. Progress of the reaction was monitored by TLC.The reaction mixture was cooled to RT and filtered through a pad ofcelite. The filtrate was concentrated under reduced pressure, and theobtained residue was dissolved in EtOAc (30 mL). The organic layer waswashed with water and brine, was dried over anhydrous Na₂SO₄, and wasconcentrated under reduced pressure. The crude compound was purified bycolumn chromatography to afford coupled product (0.21 g, 0.49 mmol, 71%)as solid. ¹H NMR (500 MHz, CDCl₃): δ 8.90 (s, 1H), 7.95 (dd, J=8.5, 2.5Hz, 1H), 7.77 (d, J=8.0 Hz, 2H), 7.71 (d, J=8.0 Hz, 2H), 7.60 (d, J=8.5Hz, 1H), 7.45-7.40 (m, 1H), 6.85 (app t, 1H), 6.75 (app t, 1H), 3.48 (d,J=5.0 Hz, 1H), 3.00 (app s, 1H). MS(ESI): m/z 428 [M⁺+1]. To a stirredsolution of coupled product (0.42 g, 0.98 mmol) in DMF (10 mL) was addedK₂CO₃ (67 mg, 0.49 mmol) followed by 1H-tetrazole (68 mg, 0.98 mmol) atRT under an inert atmosphere. The reaction mixture was stirred for 5 hat 80° C. The volatiles were removed under reduced pressure and theobtained residue was dissolved in EtOAc (30 mL). The organic layer waswashed with water and brine, was dried over anhydrous Na₂SO₄, and wasconcentrated under reduced pressure. The crude compound was purified bycolumn chromatography to afford 2 (0.14 g, 0.28 mmol, 29%) as whitesolid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.73 (s, 1H), 8.01 (dd,J=8.0, 2.0 Hz, 1H), 7.78 (d, J=8.5 Hz, 2H), 7.72-7.67 (m, 3H), 7.49 (s,1H), 7.44-7.37 (m, 1H), 6.81-6.76 (m, 1H), 6.71-6.65 (m, 1H), 5.57 (d,J=14.0 Hz, 1H), 5.19 (d, J=14.0 Hz, 1H). HPLC: 97.3%. MS(ESI): m/z 498[M⁺+1].

Chiral Preparative High Performance Liquid Chromatography (HPLC) ofEnantiomers:

The enantiomers of 2 (150 mg, 0.3 mmol) were separated by normal-phasepreparative high performance liquid chromatography (Chiralpak IC,250×21.2 mm, 5μ; using (A) n-hexane-(B) isopropyl alcohol (IPA) (A:B60:40) as a mobile phase; flow rate: 11 mL/min) to obtain 2(+) (40 mg)and 2(−) (40 mg).

Analytical Data for 2 (+):

HPLC: 100%.

Chiral HPLC: R_(t)=22.7 min (Chiralpak IC, 250×4.6 mm, 5μ; mobile phase(A) n-Hexane-(B) IPA A:B 60:40; flow rate: 1.00 mL/min)

Optical rotation [α]_(D) ²⁵: +18° (C.=0.1% in methyl alcohol (MeOH)).

Example 3

3-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)benzonitrile(3)

Compound 3 was prepared using the conditions employed for 1. 0.020 g of3 was isolated as a tan solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H),8.71 (s, 1H), 7.99 (dd, J=8.0, 2.0 Hz, 1H), 7.84 (s, 1H), 7.80-7.76 (m,2H), 7.72 (d, J=8.0 Hz, 1H), 7.65 (t, J=7.5 Hz, 1H), 7.43-7.38 (m, 2H),6.81-6.76 (m, 1H), 6.72-6.68 (m, 1H), 5.54 (d, J=14.5 Hz, 1H), 5.20 (d,J=14.5 Hz, 1H). HPLC: 93.95%. MS (ESI): m/z 455 [M⁺+1].

Example 4

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-isopropoxyphenyl)pyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(4)

Compound 4 was prepared using the conditions employed for 1: 0.029 g of4 was isolated as a white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s,1H), 8.71 (s, 1H), 7.94 (dd, J=8.5, 2.5 Hz, 1H), 7.82 (s, 1H), 7.61 (d,J=8.0 Hz, 1H), 7.50-7.47 (m, 2H), 7.40-7.35 (m, 1H), 7.01-6.98 (m, 2H),6.79-6.74 (m, 1H), 6.68-6.64 (m, 1H), 5.61 (d, J=14.0 Hz, 1H), 5.10 (d,J=14.0 Hz, 1H), 4.64-4.59 (m, 1H), 1.37 (d, J=6.0 Hz, 6H). HPLC: 99.1%.MS (ESI): m/z 488 [M⁺+1].

Example 5

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenyl)pyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(5)

Compound 5 was prepared using the conditions employed for 1: 0.033 g of5 was isolated as a white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s,1H), 8.69 (s, 1H), 7.95 (dd, J=8.0, 2.0 Hz, 1H), 7.66 (d, J=8.5 Hz, 2H),7.55-7.52 (m, 2H), 7.42-7.37 (m, 1H), 7.22-7.19 (m, 2H), 6.80-6.75 (m,1H), 6.70-6.66 (m, 1H), 5.58 (d, J=14.5 Hz, 1H), 5.15 (d, J=14.5 Hz,1H). HPLC: 99.7%. MS (ESI): m/z 448 [M⁺+1].

Example 6

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(3-(trifluoromethoxy)phenyl)pyridin-2-yl)propan-2-ol(6)

Compound 6 was prepared using the conditions employed for 1: 0.028 g of6 was isolated as a yellow solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s,1H), 8.73 (s, 1H), 7.98 (dd, J=8.0, 2.2 Hz, 1H), 7.69 (d, J=8.5 Hz, 1H),7.57-7.49 (m, 3H), 7.41-7.33 (m, 3H), 6.80-6.75 (m, 1H), 6.70-6.66 (m,1H), 5.59 (d, J=14.5 Hz, 1H), 5.16 (d, J=14.5 Hz, 1H). HPLC: 97.2%. MS(ESI): m/z 514 [M⁺+1].

Example 7

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyridin-2-yl)propan-2-ol (7)

To a stirred solution of bromo epoxide C (0.5 g, 1.38 mmol) in THF (30mL) and water (14 mL) were added (4-(trifluoromethoxy)phenyl)boronicacid (0.22 g, 1.1 mmol), Na₂CO₃ (0.32 g, 3.1 mmol) and Pd(dppf)₂Cl₂(0.28 g, 0.34 mmol) at RT under an inert atmosphere. After purging withargon for a period of 30 min, the reaction mixture was heated to 75° C.and stirring was continued for 4 h. Progress of the reaction wasmonitored by TLC. The reaction mixture was cooled to RT and filteredthrough a pad of celite. The filtrate was concentrated under reducedpressure; the obtained residue was dissolved in EtOAc (30 mL). Theorganic layer was washed with water and brine, was dried over anhydrousNa₂SO₄, and was concentrated under reduced pressure. The crude compoundwas purified by column chromatography to afford the coupled product(0.45 g, 1.0 mmol, 73%) as solid. ¹H NMR (200 MHz, CDCl₃): δ 8.87 (s,1H), 7.90 (dd, J=8.2, 2.2 Hz, 1H), 7.66-7.54 (m, 3H), 7.49-7.34 (m, 3H),6.90-6.70 (m, 2H), 3.49 (d, J=5.0 Hz, 1H), 3.02-2.95 (m, 1H). MS(ESI):m/z 444 [M⁺+1].

To a stirred solution of the coupled product (0.45 g, 1.0 mmol) in DMF(10 mL) was added K₂CO₃ (70 mg, 0.5 mmol) followed by 1H-tetrazole (70mg, 1.0 mmol) at RT under an inert atmosphere. The reaction mixture wasstirred for 4 h at 80° C. The volatiles were removed under reducedpressure, and the obtained residue was dissolved in water (15 mL) andwas extracted with EtOAc (2×20 mL). The combined organic layers werewashed with water and brine and were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude compound was purified bycolumn chromatography to afford 7 (0.19 g, 0.37 mmol, 36%) as whitesolid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.70 (s, 1H), 7.97 (dd,J=8.0, 2.0 Hz, 1H), 7.68 (d, J=8.5 Hz, 1H), 7.60-7.56 (m, 3H), 7.43-7.36(m, 3H), 6.80-6.76 (m, 1H), 6.70-6.67 (m, 1H), 5.57 (d, J=14.5 Hz, 1H),5.17 (d, J=14.5 Hz, 1H). HPLC: 98.3%. MS(ESI): m/z 513.9 [M⁺+1].

Chiral Preparative HPLC of Enantiomers:

The enantiomers of 7 (17.8 g, 34.6 mmol) were separated by normal-phasepreparative HPLC (Chiralpak AD-H, 250×21.2 mm, 5μ; using (A)n-hexane-(B) IPA (A:B 70:30) as a mobile phase; flow rate: 15 mL/min) toobtain 7(+) (6.0 g) and 7(−) (5.8 g).

Analytical Data for 7 (+):

HPLC: 99.8%.

Chiral HPLC: R_(t)=9.88 min (Chiralpak AD-H, 250×4.6 mm, 5μ; mobilephase (A) n-Hexane-(B) IPA A:B 70:30; flow rate: 1.00 mL/min)

Optical rotation [α]_(D) ²⁵: +19° (C.=0.1% in MeOH).

Example 8

1-(5-(3-Chlorophenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(8)

Compound 8 was prepared using the conditions employed for 1: 0.028 g of8 was isolated as a white solid. ¹H NMR (500 MHz, CDCl₃): δ□8.76 (s,1H), 8.72 (s, 1H), 7.97 (dd, J=8.5, 2.2 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H),7.56-7.54 (m, 2H), 7.46-7.43 (m, 3H), 7.40-7.35 (m, 1H), 6.80-6.75 (m,1H), 6.70-6.66 (m, 1H), 5.59 (d, J=14.5 Hz, 1H), 5.16 (d, J=14.5 Hz,1H). HPLC: 98.79%. MS (ESI): m/z 463.9 [M+]⁺.

Example 9

1-(5-(4-Chlorophenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(9)

Compound 9 was prepared using the conditions employed for 1: 0.027 g of9 was isolated as a white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.75 (s,1H), 8.70 (s, 1H), 7.96 (d, J=8.5 Hz, 1H), 7.66 (d, J=8.5 Hz, 1H), 7.60(s, 1H), 7.49 (s, 4H), 7.42-7.37 (m, 1H), 6.79-6.76 (m, 1H), 6.70-6.67(m, 1H), 5.58 (d, J=14.5 Hz, 1H), 5.16 (d, J=14.5 Hz, 1H). HPLC: 99.07%.MS (ESI): m/z 463.9 [M+]⁺.

Chiral Preparative HPLC of Enantiomers:

The enantiomers of 9 (200 mg, 0.4 mmol) were separated by normal-phasepreparative HPLC (Chiralpak IC, 250×21.1 mm, 5μ; using (A) n-hexane-(B)ethyl alcohol (A:B 75:25) as a mobile phase; flow rate: 15 mL/min) toobtain 9(+) (62 mg) and 9(−) (55 mg).

Analytical Data for 9 (+):

HPLC: 100%

Chiral HPLC: R_(t)=15.3 min (Chiralpak IC, 250×4.6 mm, 5μ; mobile phase(A) n-Hexane-(B)ethyl alcohol A:B 75:25; flow rate: 1.00 mL/min)

Optical rotation [α]_(D) ²⁵: +26.5° (C.=0.1% in MeOH).

Example 10

2-(2,4-Difluorophenyl)-1-(5-(2,5-difluorophenyl)pyridin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(10)

Compound 10 was prepared using the conditions employed for 1: 0.022 g of10 was isolated as a yellow solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s,1H), 8.70 (s, 1H), 7.98 (d, J=8.0 Hz, 1H), 7.69 (d, J=8.0 Hz, 1H), 7.49(s, 1H), 7.41-7.36 (m, 1H), 7.20-7.11 (m, 3H), 6.79-6.75 (m, 1H),6.70-6.67 (m, 1H), 5.60 (d, J=14.5 Hz, 1H), 5.16 (d, J=14.5 Hz, 1H).HPLC: 98.68%. MS (ESI): m/z 466 [M+]⁺.

Example 11

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol(11)

Compound 11 was prepared using the conditions employed for 1: 0.33 g of11 was isolated as a solid. The precursor1-bromo-4-(2,2,2-trifluoroethoxy)benzene was prepared as described belowin one step.

¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.70 (s, 1H), 7.95 (d, J=8.0Hz, 1H), 7.70 (s, 1H), 7.64 (d, J=8.5 Hz, 1H), 7.54 (d, J=8.5 Hz, 2H),7.42-7.37 (m, 1H), 7.08 (d, J=8.5 Hz, 2H), 6.79-6.75 (m, 1H), 6.69-6.66(m, 1H), 5.58 (d, J=14.0 Hz, 1H), 5.14 (d, J=14.0 Hz, 1H), 4.44-4.39 (m,2H). HPLC: 99.1%. MS (ESI): m/z 528 [M⁺+1].

Chiral Preparative HPLC of Enantiomers:

The enantiomers of 11 (330 mg, 0.626 mmol) were separated bynormal-phase preparative HPLC (Chiralpak IC, 250×21.1 mm, 5μ; using (A)n-hexane-(B) IPA (A:B 65:35) as a mobile phase; Flow rate: 15 mL/min) toobtain 11(+) (126.3 mg) and 11(−) (112.7 mg).

Analytical Data for 11 (+):

HPLC: 99.8%

Chiral HPLC: R_(t)=13.40 min (Chiralpak IA, 250×4.6 mm, 5μ; mobile phase(A) n-Hexane-(B) IPA A:B 65:35; flow rate: 1.00 mL/min)

Optical rotation [α]_(D): +24° (C.=0.1% in MeOH). Optical rotation[α]_(D): +24° (C.=0.1% in MeOH).

1-Bromo-4-(2,2,2-trifluoroethoxy)benzene

To a stirred solution of trifluoroethyl tosylate (1.5 g, 5.8 mmol) inDMF (20 mL) was added K₂CO₃ (4 g, 29.4 mmol) followed by p-bromo phenol(1.1 g, 6.46 mmol) at RT under an inert atmosphere. The reaction mixturewas stirred at 120° C. for 6 h. The volatiles were evaporated underreduced pressure; the residue was diluted with water (5 mL) andextracted with EtOAc (3×30 mL). The organic layer was washed with waterand brine, was dried over anhydrous Na₂SO₄, was filtered and wasconcentrated in vacuo. The crude compound was purified by silica gelcolumn chromatography eluting with 5% EtOAc/hexane to afford the desiredproduct (0.8 g, 3.13 mmol, 53.3%) as semi solid. ¹H NMR (200 MHz,CDCl₃): δ 7.44-7.38 (m, 2H), 6.86-6.80 (m, 2H), 4.38-4.25 (m, 2H).

Example 12

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-(2,2,3,3,3-pentafluoropropoxy)phenyl)pyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(12)

To a stirred solution of trifluorethanol (10 g, 0.06 moles (mol)) in dryCH₂Cl₂ (100 mL) was added N,N-diisopropylethylamine (DIPEA; 29 mL, 0.16mol) at RT, and the reaction mixture was cooled to −78° C. Triflicanhydride (13.5 mL, 0.07 mol) was added dropwise to the reaction mixtureat −78° C. After being stirred for 30 min, the reaction mixture waswarmed to −30° C. and stirring was continued for another 30 min. Thereaction mixture was quenched with water (200 mL) and extracted withCH₂C12 (2×300 mL). The combined organic layers were washed with 1 Nhydrochloric acid (HCl) and water, dried over anhydrous Na₂SO₄, andfiltered. To a stirred solution of 4-bromophenol (4 g, 0.02 mol) andcesium carbonate (Cs₂CO₃; 15 g, 0.04 mol) in DMF (100 mL) was added theCH₂Cl₂ layer from above (H) at RT. The mixture was stirred for 16 h. Theprogress of the reaction was monitored by TLC. The reaction mixture wasdiluted with water and extracted with CH₂C12 (2×250 mL). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure. The obtained crude material waspurified by column chromatography (SiO₂, 60-120 mesh) to afford compoundF (3.5 g, 11.5 mmol, 50%) as a liquid. ¹H NMR (200 MHz, CDCl₃): δ7.46-7.38 (m, 2H), 6.87-6.79 (m, 2H), 4.45-4.32 (m, 2H).

To a stirred solution of n-BuLi (21 mL, 33.13 mmol, 1.5 M in hexane) indry ether (250 mL) was added a solution of compound C (8 g, 22.09 mmol)in ether (50 mL) at −78° C. After being stirred for 30 min, trimethylborate (5 mL, 44.19 mmol) was added to the reaction mixture at −78° C.and the stirring was continued for another 10 min. The reaction mixturewas allowed to warm to room temperature and was stirred for 30 min. Thereaction mixture was quenched with acetic acid (40 mL), was diluted withwater (120 mL) and was stirred for 1 h at RT. The reaction mixture wasbrought to pH˜12 by the addition of 2 N sodium hydroxide (NaOH), theorganic layer was separated and the aqueous layer was brought to pH-6using 1 N HCl. The aqueous layer was extracted with CH₂Cl₂ (2×500 mL).The combined organic layers were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford compound E (7 g, 21.4mmol, 97%) as a brown white solid. ¹H NMR (500 MHz, CD₃OD): δ 8.81 (s,1H), 8.15 (d, J=7.5 Hz, 1H), 7.47 (d, J=8 Hz, 1H), 7.36-7.35 (m, 1H),6.93-6.87 (m, 2H), 3.42 (d, J=5.5 Hz, 1H), 2.99-2.98 (m, 1H). MS (ESI):m/z 328.1 [M⁺+1]. A mixture of boronic acid E (3.5 g, 10.7 mmol),compound F (3.3 g, 10.7 mol) and K₂CO₃ (4.5 g, 32.1 mmol) in THF/H₂O(175 mL, 4:1) was degassed for 30 min. Pd (dppf)₂ Cl₂ (0.7 g, 1.07 mmol)was added to the reaction mixture under an an inert atmosphere, and theresulting mixture was stirred at 70° C. for 2 h. The reaction mixturewas allowed to cool to room temperature and the volatiles were removedunder reduced pressure. The obtained crude material was purified bycolumn chromatography (SiO₂, 60-120 mesh; eluent: 15-55% EtOAc/hexanes)to afford compound G (2.3 g, 4.53 mmol, 43%) as an off-white solid. ¹HNMR (200 MHz, CDCl₃): δ 8.83 (d, J=2.2 Hz, 1H), 7.90 (dd, J=2.2, 8.0 Hz,1H), 7.61-7.48 (m, 3H), 7.43-7.36 (m, 1H), 7.29 (d, J=8.8 Hz, 2H),7.10-7.04 (m, 2H), 6.89-6.70 (m, 2H), 4.48 (q, J=12.4 Hz, 2H), 3.45 (d,J=5.0 Hz, 1H), 3.01-2.98 (m, 1H).

To a stirred solution of compound G (10.5 g, 20.7 mmol) in DMF (150 mL)was added K₂CO₃ (3.4 g, 20.7 mmol) followed by 1H-tetrazole (2.6 g, 37.1mmol) at RT. The reaction mixture was heated to 70° C. for 16 h.Progress of the reaction was monitored by TLC. The reaction mixture wasallowed to cool to room temperature and diluted with water (300 mL). Theaqueous layer was extracted with EtOAc (3×300 mL). The combined organiclayer was dried over anhydrous Na₂SO₄ and concentrated in vacuo. Thecrude compound was purified by column chromatography (SiO₂, 60-120 mesh;eluent: 15-55% EtOAc/hexanes) to afford 12 (6 g, 10.38 mmol, 50.4%) as awhite solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.70 (s, 1H), 7.95(d, J=8.0 Hz, 1H), 7.70 (s, 1H), 7.64 (d, J=8.5 Hz, 1H), 7.54 (d, J=8.5Hz, 2H), 7.42-7.37 (m, 1H), 7.08 (d, J=8.5 Hz, 2H), 6.79-6.75 (m, 1H),6.69-6.66 (m, 1H), 5.58 (d, J=14.0 Hz, 1H), 5.14 (d, J=14.0 Hz, 1H),4.48 (t, J=12.0 Hz, 2H). MS (ESI): m/z 578.1 [M⁺+1].

Chiral Preparative HPLC of Enantiomers:

The enantiomers of 12 (6 g, 10.3 mmol) were separated by normal-phasepreparative HPLC (Chiralpak IA, 250×21.2 mm, 5μ; using (A)n-hexane-(B)ethyl alcohol (A:B 80:20) as a mobile phase; flow rate: 12mL/min) to obtain 12(+) (2.1 g) and 12(−) (2.0 g).

Analytical Data for 12 (+):

¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.70 (s, 1H), 7.95 (d, J=8.0Hz, 1H), 7.70 (s, 1H), 7.64 (d, J=8.5 Hz, 1H), 7.54 (d, J=8.5 Hz, 2H),7.42-7.37 (m, 1H), 7.08 (d, J=8.5 Hz, 2H), 6.79-6.75 (m, 1H), 6.69-6.66(m, 1H), 5.58 (d, J=14.0 Hz, 1H), 5.14 (d, J=14.0 Hz, 1H), 4.48 (t,J=12.0 Hz, 2H). HPLC: 98.1%. MS (ESI): m/z 578.1 [M⁺+1].

Chiral HPLC: R_(t)=14.12 min (Chiralpak IA, 250×4.6 mm, 5μ; mobile phase(A) n-Hexane-(B)ethyl alcohol A:B 80:20); flow rate: 1.00 mL/min).

Optical rotation [α]_(D) ²⁵: +22.3° (C.=0.1% w/v in MeOH).

Example 13

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl3-aminopropanoate hydrochloride (13)

To a mixture of Boc-βalanine (N-Boc-β-Ala-OH; 1 g, 5.29 mmol) andN-hydroxysuccinimide (0.9 g, 7.82 mmol) in DMF (10 mL) were added1-hydroxybenzotriazole hydrate (HOBt.xH₂O; 0.7 g, 5.25 mmol) and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl; 1g, 5.23 mmol) at 5° C. The reaction mixture was warmed to RT and stirredfor 16 h. The progress of the reaction was monitored by TLC. Thereaction was quenched with water and the mixture was extracted withEtOAc (2×150 mL). The combined organic layers were washed with water(3×100 mL) and brine (150 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude compound was trituratedwith ether (2×25 mL) to afford N-Boc-β-Ala-OSu (1.1 g, crude) as a whitesolid. ¹H NMR (500 MHz, CDCl₃): δ 5.10 (br s, 1H), 3.52 (q, J=6.0 Hz,2H), 2.85-2.82 (m, 6H), 1.31 (s, 9H).

To a suspension of 11-(+) (0.2 g, 0.38 mmol) in dry THF (20 mL) wasadded sodium hydride (NaH; 0.02 g, 1.17 mmol) at 0° C., and the mixturewas stirred for 30 min at RT. N-Boc-β-Ala-OSu (0.21 g, 0.70 mmol) wasadded to the reaction mixture and the stirring was continued for another16 h at RT. The progress of the reaction was monitored by TLC. Thereaction mixture was quenched with ice cold water and was extracted withEtOAc (2×50 mL). The combined organic layers were dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to afford crude product,which upon separation by preparative TLC (SiO₂, 60-120 mesh; eluent:15-55% EtOAc/hexanes) afforded compound I (38 mg, 0.06 mmol, 15%). NMR(500 MHz, CDCl₃): δ 9.27 (s, 1H), 8.92 (s, 1H), 7.80 (dd, J=1.5, 8.0 Hz,1H), 7.58 (d, J=8.5 Hz, 2H), 7.14-7.13 (m, 1H), 7.09 (d, J=8.5 Hz, 2H),7.04 (d, J=8.0 Hz, 1H), 6.89 (t, J=7.0 Hz, 1H), 6.71-6.66 (m, 1H), 6.09(dd, J=2.5, 15.0 Hz, 1H), 5.73 (dd, J=2.5, 15.0 Hz, 1H), 5.23 (br s,1H), 4.45-4.40 (m, 2H), 3.46 (br s, 2H), 2.82-2.69 (m, 2H), 1.28 (s,9H). MS (ESI): m/z 699.3 [M⁺+1].

To a stirred solution of compound I (0.03 g, 0.05 mmol) in 1,4-dioxane(2 mL) was added 4 M HCl solution in 1,4-dioxane (1 mL) at 5° C., andthe mixture was stirred for 4 h at RT. The progress of the reaction wasmonitored by TLC. The volatiles were evaporated under reduced pressure.The obtained crude was triturated with diethyl ether (2×25 mL) to afford13 (0.018 g, 0.02 mmol, 55%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆): δ 9.67 (s, 1H), 9.04 (s, 1H), 8.13 (dd, J=1.5, 8.0 Hz, 1H),7.88 (s, 2H), 7.78 (d, J=8.5 Hz, 2H), 7.38-7.36 (m, 1H), 7.27-7.24 (m,1H), 7.24 (d, J=8.0 Hz, 1H), 7.17 (d, J=8.0 Hz, 1H), 6.15 (d, J=15.5 Hz,1H), 5.54 (d, J=15.5 Hz, 1H), 4.87 (q, J=8.5 Hz, 2H), 3.06 (d, J=5.5 Hz,2H), 2.93-2.83 (m, 2H). HPLC: 93.64%. MS (ESI): m/z 599.4 [M⁺+1].

Example 14

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl2-aminoacetate hydrochloride (14)

To a suspension of 11-(+) (0.1 g, 0.18 mmol) in dry THF (30 mL) wasadded NaH (0.01 g, 0.41 mmol) at 5° C. and the mixture was stirred for40 min at RT. Boc-Glycine N-hydroxysuccinimide ester (N-Boc-Gly-OSu; 0.1g, 0.37 mmol) was added to the reaction mixture, and the stirring wascontinued for another 16 h at RT. The progress of the reaction wasmonitored by TLC. The reaction was quenched with ice cold water andextracted with EtOAc (2×50 mL). The combined organic layers were driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to affordcrude product, which upon separation by preparative TLC (SiO₂, 60-120mesh; eluent: 15-55% EtOAc/hexanes) afforded compound J (29 mg, 0.04mmol, 24%). ¹H NMR (500 MHz, DMSO-d₆): δ 9.34 (s, 1H), 8.92 (s, 1H),7.80 (d, J=7.0 Hz, 1H), 7.59-7.54 (m, 2H), 7.44-7.42 (m, 1H), 7.10-7.03(m, 3H), 6.94-6.91 (m, 1H), 6.64 (t, J=10.0 Hz, 1H), 6.12 (dd, J=2.5,15.0 Hz, 1H), 5.69 (dd, J=3.5, 15.0 Hz, 1H), 5.10 (d, J=6.0 Hz, 1H),4.43 (q, J=8.5 Hz, 2H), 4.21-4.16 (m, 1H), 3.95 (dd, J=5.0, 18.0 Hz,1H), 1.45 (s, 9H). MS (ESI): m/z 685.3 [M⁺+1].

To a stirred solution of compound J (0.02 g, 0.04 mmol) in 1,4-dioxane(2 mL) was added a 4 M HCl solution in 1,4-dioxane (1 mL) dropwise at 5°C. The mixture was stirred for 4 h at RT. The progress of the reactionwas monitored by TLC. The volatiles were evaporated under reducedpressure. The obtained crude product was triturated with diethyl ether(3×25 mL) to afford 14 (14 mg, 0.02 mmol, 60%) as a white solid. ¹H NMR(500 MHz, DMSO-d₆): δ 9.68 (s, 1H), 9.04 (s, 1H), 8.45-8.43 (m, 2H),8.14 (d, J=8.5 Hz, 2H), 7.79 (d, J=9.0 Hz, 2H), 7.45-7.44 (m, 1H),7.29-7.27 (m, 1H), 7.24-7.23 (m, 3H), 7.14-7.10 (m, 1H), 6.18 (d, J=16.0Hz, 1H), 5.57 (d, J=15.0 Hz, 1H), 4.87 (q, J=8.5 Hz, 2H), 4.16 (d,J=18.0 Hz, 1H), 3.94 (d, J=18.5 Hz, 1H). HPLC: 93.54%. MS (ESI): m/z 585[M⁺+1].

Example 15

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-pyrazol-3-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyridin-2-yl)propan-2-ol (15)

To a suspension of copper powder (27 g, 0.42 mol) in DMSO (300 mL) wasadded ethyl α-bromo-difluoroacetate (27 mL, 0.21 mol) and the mixturewas stirred for 1 h at RT. 2,5-Dibromopyridine (25 g, 0.10 mol) was thenadded and stirring was continued for another 15 h at RT. The progress ofthe reaction was monitored by TLC. The reaction was quenched withsaturated NH₄Cl solution (200 mL) and extracted with CH₂C12 (3×250 mL).The combined organic layers were washed with water and brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to afford crudeproduct, which upon distillation under reduced pressure affordedcompound K (19 g, 67.8 mmol, 64%) as a pale yellow oil. ¹H NMR (500 MHz,CDCl₃): δ 8.71 (s, 1H), 8.00 (d, J=9.0 Hz, 1H), 7.62 (d, J=9.0 Hz, 1H),4.42-4.35 (m, 2H), 1.39-1.31 (m, 3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (7.6 mL, 67.8 mmol)in diethyl ether (100 mL) was added n-BuLi (42 mL, 67.85 mmol, 1.6 M inhexane) at −78° C. After being stirred for 45 min at −78° C., a solutionof ester K (19 g, 67.8 mmol) in diethyl ether (100 mL) was added to thereaction mixture and the stirring was continued for another 1 h at −78°C. under an inert atmosphere. The reaction mixture was warmed to RT andwas stirred for another 3 h. The progress of the reaction was monitoredby TLC. The reaction was quenched with saturated NH₄Cl solution (200 mL)and the reaction mixture was extracted with EtOAc (3×200 mL). Thecombined organic layers were washed with water and brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The crudecompound was purified by column chromatography (SiO₂, 100-200 mesh)eluting with 2% EtOAc/hexane to afford ketone L (13 g, 37.3 mmol, 55%)as yellow liquid. ¹H NMR (500 MHz, CDCl₃): δ 8.62 (s, 1H), 8.08-8.04 (m,2H), 7.72 (d, J=8.5 Hz, 1H), 7.05-6.95 (m, 1H), 6.88-6.78 (m, 1H). MS(ESI): m/z 347[M⁺+1], 349 [(M⁺+2].

To a stirred solution of ketone L (1.0 g, 2.87 mmol) in THF (30 mL) andwater (10 mL) were added (4-(trifluoro methoxy)phenyl boronic acid (591mg, 2.87 mmol), sodium bicarbonate (NaHCO₃; 782 mg, 7.18 mmol) andPd(dppf)₂Cl₂ (586 mg, 0.718 mmol) at RT under an inert atmosphere. Afterpurging with argon for a period of 30 min, the reaction mixture washeated to 65° C. and stirring was continued for 2 h. Progress of thereaction was monitored by TLC. The reaction mixture was cooled to RT andfiltered through a pad of celite. The filtrate was concentrated underreduced pressure, and the obtained residue was dissolved in EtOAc (2×50mL). The organic layer was washed with water, brine and dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The crudecompound was purified by column chromatography (SiO₂, 100-200 mesh;eluent: 15-55% EtOAc/hexanes) to afford M (980 mg, 2.28 mmol, 79%) aslight yellow sticky solid. ¹H NMR (200 MHz, CDCl₃): δ 8.77 (s, 1H),8.12-8.03 (m, 2H), 7.90 (d, J=8.4 Hz, 1H), 7.63-7.57 (m, 2H), 7.35 (d,J=8.2 Hz, 2H), 7.05-6.96 (m, 1H), 6.83-6.79 (m, 1H). MS(ESI): m/z 430[M⁺+1].

To a mixture of magnesium (Mg; 50 mg, 2.08 mmol) and mercuric chloride(HgCl₂; 47 mg, 0.17 mmol) in dry THF (5 mL) was added propargyl bromide(0.05 mL, 0.34 mmol) at RT under an an inert atmosphere, and the mixturewas stirred for 20 min. The reaction mixture was then cooled to −20° C.,and ketone M (150 mg, 0.348 mmol) and the remaining portion of propargylbromide (0.05 mL, 0.34 mmol) in THF (5 mL) were added. Stirring wascontinued stirring for 2 h at −20° C. The progress of the reaction wasmonitored by TLC. The reaction was quenched with a saturated NH₄Clsolution and the reaction mixture was extracted with CH₂Cl₂ (3×50 mL).The combined organic layers were washed with water and brine, dried overanhydrous Na₂SO₄ and concentrated in vacuo. The crude product waspurified by column chromatography (SiO₂, 100-200 mesh; eluent: 15-55%EtOAc/hexanes) to afford N (110 mg, 0.23 mmol, 67%) as a solid. ¹H NMR(200 MHz, CDCl₃): δ 8.86 (s, 1H), 7.96 (dd, J=8.4, 2.2 Hz, 1H),7.65-7.57 (m, 4H), 7.41 (d, J=8.2 Hz, 2H), 6.88-6.73 (m, 2H), 6.36 (brs,1H), 3.46 (dd, J=16.8, 2.2 Hz, 1H), 2.98 (dt, J=16.8, 2.6 Hz, 1H), 1.85(t, J=2.6 Hz, 1H). MS (ESI): m/z 470 [M⁺+1].

A solution of N (110 mg, 0.23 mmol) in TMSCHN₂ (1 mL, 1.15 mmol) wasstirred at 120° C. for 15 h. The volatiles were evaporated under reducedpressure and the obtained crude material was purified by columnchromatography (SiO₂, 100-200 mesh; eluent: 15-55% EtOAc/hexanes) toafford 15 (35 mg, 0.06 mmol, 29%) as an off white solid. ¹H NMR (500MHz, CDCl₃): δ 8.80 (s, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.62-7.59 (m, 3H),7.50-7.45 (m, 1H), 7.36-7.31 (m, 3H), 6.83 (br s, 1H), 6.70-6.65 (m,2H), 6.04 (s, 1H), 4.02 (d, J=15.0 Hz, 1H), 3.36 (d, J=15.0 Hz, 1H). MS(ESI): m/z 512 [M⁺+1]. HPLC: 95.6%.

Example 16

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenyl)pyridin-2-yl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol(16)

To a stirred solution of5-bromo-2-((2-(2,4-difluorophenyl)oxiran-2-yl)difluoromethyl)pyridine(C; 1.0 g, 2.7 mmol) in THF:H₂O (20 mL, 4:1 mixture) was added(4-fluorophenyl)boronic acid (378 mg, 2.7 mmol) followed by K₂CO₃ (1.1g, 8.1 mmol) at RT, and the mixture was degassed by purging with inertgas for 45 min. To the resulting reaction mixture was added Pd(dppf)₂Cl₂(197 mg, 0.27 mmol), and the reaction mixture was further degassed for20 min at RT. The reaction mixture was then heated to 60° C. and stirredfor 4 h. After complete consumption of the starting material (by TLC),the reaction mixture was cooled to RT, diluted with water and theorganic layer separated. The aqueous layer with extracted with EtOAc(2×20 mL). The combined organic extracts were dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to obtain the crudeproduct. The crude material was purified by silica gel columnchromatography (eluent: 20% EtOAc/hexane) to afford 0 (0.9 g, 2.38 mmol,86%) as colorless semi-solid. ¹H NMR (200 MHz, CDCl₃): δ 8.85 (d, J=2.0Hz, 1H), 7.89 (dd, J=8.2, 2.4 Hz, 1H), 7.62-7.36 (m, 4H), 7.24-7.19 (m,2H), 6.90-6.70 (m, 2H), 3.48 (d, J=4.8 Hz, 1H), 3.02-2.98 (m, 1H).

To a stirred solution of compound O (0.3 g, 0.79 mmol) in DMF (3 mL) wasadded K₂CO₃ (109 mg, 0.79 mmol) followed by 1,2,4-triazole (81 mg, 1.18mmol) at RT under an inert atmosphere. The reaction mixture was thenheated to 60° C. and stirred for 16 h. After complete consumption of thestarting material (by TLC), the reaction mixture was diluted with waterand extracted with EtOAc (3×15 mL). The combined organic extracts weredried over anhydrous Na₂SO₄ and concentrated under reduced pressure toobtain the crude product. The crude material was purified by silica gelcolumn chromatography (eluent: 40% EtOAc/hexane) to afford 16 (250 mg,0.56 mmol, 72.6%) as an off-white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.72(s, 1H), 8.16 (s, 1H), 7.92 (d, J=8.5 Hz, 1H), 7.69 (s, 1H), 7.62 (d,J=8.5 Hz, 1H), 7.56-7.47 (m, 3H), 7.22-7.18 (m, 2H), 6.77-6.71 (m, 3H),5.38 (d, J=14.0 Hz, 1H), 4.90 (d, J=14.0 Hz, 1H). MS (ESI): m/z 447[M⁺+1]. HPLC: 98.36%.

Example 17

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-1,2,4-triazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol(17)

To a stirred solution of epoxy bromide C (190 mg, 0.52 mmol) in THF:H₂O(40 mL, 4:1 mixture) was added (4-(2,2,2-trifluoroethoxy)phenyl)boronicacid (174 mg, 0.57 mmol) followed by K₂CO₃ (215 mg, 1.56 mmol) at RT,and the mixture was degassed by purging with inert gas for 30 min. Tothe resulting reaction mixture was added Pd(dppf)₂Cl₂ (20 mg, 0.027mmol), and the mixture was further degassed for 20 min at RT. Thereaction mixture was then heated to 70° C. and stirred for 2 h. Progressof the reaction was monitored by TLC. The reaction mixture was cooled toRT, diluted with EtOAc (20 mL) and filtered through a celite pad. Thecollected filtrate was washed with water (2×50 mL). The separatedorganic layer was dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude product. The crude material waspurified by silica gel column chromatography (eluent: 15% EtOAc/hexane)to afford P (0.2 g, 0.43 mmol, 84%) as an off-white solid. ¹H NMR (200MHz, CDCl₃): δ 8.85 (d, J=2.2 Hz, 1H), 7.89 (dd, J=8.2, 2.2 Hz, 1H),7.59-7.51 (m, 3H), 7.48-7.36 (m, 1H), 7.08 (dd, J=7.0, 2.2 Hz, 2H),6.89-6.70 (m, 2H), 4.42 (q, J=8.2 Hz, 2H), 3.48 (d, J=5.0 Hz, 1H),3.01-2.98 (m, 1H). MS (ESI): m/z 458 [M⁺+1].

To a stirred solution of compound P (0.2 g, 0.43 mmol) in DMF (20 mL)was added K₂CO₃ (91 mg, 0.65 mmol) followed by 1,2,4-triazole (61 mg,0.87 mmol) at RT under an inert atmosphere. The reaction mixture wasthen heated to 75° C. and stirred for 7 h. After complete consumption ofthe starting material (by TLC), the reaction mixture was cooled to RT,diluted with water and extracted with EtOAc (3×75 mL). The combinedorganic extracts were dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude product. The crude material waspurified by silica gel column chromatography (eluent: 40% EtOAc/Hexane)to afford 17 (160 mg, 0.303 mmol, 70%) as an off-white solid.

Chiral Preparative HPLC of Enantiomers

The enantiomers of 17 (100 mg, 0.18 mmol) were separated by normal-phasepreparative HPLC (Chiralpak IC, 250×19 mm, 5μ; using (A) n-hexane-(B)IPA (A:B 60:40) as a mobile phase; flow rate: 15 mL/min, λ=265 nm) toobtain desired 17-(+) (28 mg) (Fraction-II) and 17-(−) (28 mg)(Fraction-I).

Analytical Data for 17(+):

¹H NMR (500 MHz, CDCl₃): 8.72 (s, 1H), 8.16 (s, 1H), 7.92 (dd, J=8.5,2.0 Hz, 1H), 7.69 (s, 1H), 7.61 (d, J=8.0 Hz, 1H), 7.55 (d, J=8.0 Hz,2H), 7.52-7.47 (m, 1H), 7.08 (d, J=9.0 Hz, 2H), 6.77-6.70 (m, 3H), 5.38(d, J=14.5 Hz, 1H), 4.89 (d, J=14.5 Hz, 1H), 4.42 (q, J=8.0 Hz, 2H).HPLC: 99.86%. MS (ESI): m/z 527 [M⁺+1].

Chiral HPLC: 99.9% ee (R_(t)=13.9 min) (Chiralpak IC, 250×4.6 mm, 5μ;mobile phase (A) n-Hexane-(B) IPA A:B 60:40; flow rate: 1 mL/min, WL 265nm).

Optical rotation [α]_(D) ^(24.5): +13.96° (C.=0.1% w/v in MeOH).

Example 18

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-1,2,4-triazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyridin-2-yl)propan-2-ol (18)

To a stirred solution of epoxy bromide (C) (0.7 g, 1.93 mmol) in THF:H₂O(24 mL, 7:5 mixture) was added (4-(trifluoromethoxy)phenyl)boronic acid(398 mg, 1.93 mmol) followed by Pd(dppf)₂Cl₂ (394 mg, 0.48 mmol) andNa₂CO₃ (526 mg, 4.83 mmol) at RT. The mixture was degassed with argonfor 45 min and then was stirred for 3 h at reflux temperature. Aftercomplete consumption of the starting material (by TLC), the reactionmixture was cooled to RT, diluted with EtOAc (20 mL) and filteredthrough a celite bed. The collected filtrate was washed with water andbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to obtain the crude product. The crude material was purified bysilica gel column chromatography (eluent: 5% EtOAc/hexane) to affordcompound Q (0.65 g, 1.46 mmol, 76%) as an off-white solid. ¹H NMR (500MHz, CDCl₃): δ 8.86 (s, 1H), 7.91 (dd, J=7.5, 2.0 Hz, 1H), 7.62 (d,J=8.5 Hz, 2H), 7.57 (d, J=7.5 Hz, 1H), 7.44-7.40 (m, 1H), 7.36 (d, J=8.5Hz, 2H), 6.86-6.83 (m, 1H), 6.77-6.73 (m, 1H), 3.49 (d, J=5.0 Hz, 1H),3.00 (d, J=5.5 Hz, 1H). MS (ESI): m/z 444 [M⁺+1].

To a stirred solution of compound Q (0.2 g, 0.45 mmol) in DMF (5 mL) wasadded K₂CO₃ (62 mg, 0.45 mmol) followed by 1,2,4-triazole (46 mg, 0.67mmol) at RT under an inert atmosphere. The reaction mixture was thenheated to 70° C. and stirred for 3 h. After consumption of the startingmaterial (by TLC), the reaction mixture was concentrated under reducedpressure, diluted with EtOAc (20 mL), and washed with water and brine.The organic layer was dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude product. The crude material waspurified by silica gel column chromatography (eluent: 30% EtOAc/hexane)to afford 18 (0.15 g, 0.29 mmol, 64.9%) as an off-white solid. ¹H NMR(500 MHz, CDCl₃): δ 8.74 (s, 1H), 8.16 (s, 1H), 7.94 (dd, J=8.0, 2.0 Hz,1H), 7.70 (s, 1H), 7.64 (d, J=8.5 Hz, 1H), 7.60 (d, J=8.0 Hz, 2H),7.51-7.46 (m, 1H), 7.36 (d, J=8.5 Hz, 2H), 6.77-6.70 (m, 2H), 6.60 (s,1H), 5.39 (d, J=14.5 Hz, 1H), 4.91 (d, J=14.5 Hz, 1H). MS (ESI): m/z 513[M⁺+1]. HPLC: 98.86%.

Example 19

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-1,2,3-triazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyridin-2-yl)propan-2-ol(19)

To a stirred solution of compound Q (0.2 g, 0.45 mmol) in DMF (5 mL) wasadded K₂CO₃ (62 mg, 0.45 mmol) followed by 1,2,3-triazole (46 mg, 0.67mmol) at RT under an inert atmosphere. The reaction mixture was thenheated to 70° C. and stirred for 3 h. After consumption of the startingmaterial (by TLC), the reaction mixture was concentrated under reducedpressure, diluted with EtOAc (20 mL), and washed with water and brine.The organic layer was dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude product. The crude material waspurified by silica gel column chromatography (eluent: 30% EtOAc/Hexane)to afford 19 (0.1 g, 0.19 mmol, 43%) as an off-white solid. ¹H NMR (500MHz, CDCl₃): δ 8.71 (s, 1H), 7.95 (d, J=8.0 Hz, 1H), 7.68 (s, 1H), 7.67(d, J=6.0 Hz, 1H) 7.59 (d, J=8.5 Hz, 2H), 7.51 (s, 1H), 7.49-7.45 (m,1H), 7.36 (d, J=8.5 Hz, 2H), 6.77-6.69 (m, 3H), 5.55 (d, J=14.5 Hz, 1H),5.12 (d, J=14.5 Hz, 1H). MS (ESI): m/z 513 [M⁺+1]. HPLC: 98.99%.

Example 20

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol(20)

Compound 20 was prepared using the same conditions as compound 1 from Pand tetrazole (0.020 g). ¹H NMR (500 MHz, CDCl₃): δ 8.74 (s, 1H), 8.31(s, 1H), 7.95 (dd, J=8.0, 2.0 Hz, 1H), 7.66 (d, J=8.0 Hz, 1H), 7.55 (d,J=9.0 Hz, 2H), 7.48-7.43 (m, 1H), 7.08 (d, J=9.0 Hz, 2H), 7.00 (s, 1H),6.84-6.69 (m, 2H), 5.83 (d, J=14.0 Hz, 1H), 5.41 (d, J=14.0 Hz, 1H),4.42 (q, J=8.5 Hz, 2H). MS (ESI): m/z 528 [M⁺+1]. HPLC: 94.47%.

Example 21

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(3-(fluorophenyl)pyridin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol(21)

Compound 21 was prepared using the same conditions as compound 1 (0.017g). ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.32 (s, 1H), 7.98 (d,J=8.0 Hz, 1H), 7.68 (dd, J=8.5, 4.0 Hz, 1H), 7.51-7.42 (m, 2H), 7.36 (d,J=8.0 Hz, 1H), 7.29-7.28 (m, 1H), 7.18-7.15 (m, 1H), 6.84-6.79 (m, 2H),6.73-6.69 (m, 1H), 5.84 (d, J=14.0 Hz, 1H), 5.42 (d, J=14.0 Hz, 1H). MS(ESI): m/z 448.1 [M⁺+1]. HPLC: 98.60%.

Example 222-(2,4-Difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)-1-(5-(4-(trifluoromethylphenyl)pyridin-2-yl)propan-2-ol (22)

Compound 22 was prepared using the same conditions as compound 1 (0.020g). ¹H NMR (500 MHz, CDCl₃): δ 8.78 (s, 1H), 8.32 (s, 1H), 8.02 (dd,J=8.0, 2.0 Hz, 1H), 7.78 (d, J=8.5 Hz, 2H), 7.72-7.68 (m, 3H), 7.48-7.43(m, 1H), 6.84-6.79 (m, 1H), 6.73-6.71 (m, 1H), 6.69 (s, 1H), 5.85 (d,J=14.0 Hz, 1H), 5.42 (d, J=14.0 Hz, 1H). MS (ESI): m/z 498.0 [M⁺+1].HPLC: 97.72%.

Example 23

4-(6-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)pyridin-3-yl)phenol(23)

Compound 23 was prepared using the same conditions as compound 1 (0.0109g). ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.69 (s, 1H), 7.94 (dd,J=8.5, 2.5 Hz, 1H), 7.80 (s, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.45 (d, J=8.5Hz, 2H), 7.41-7.36 (m, 1H), 6.96 (d, J=8.5 Hz, 2H), 6.79-6.75 (m, 1H),6.69-6.65 (m, 1H), 5.60 (d, J=14.0 Hz, 1H), 5.17 (br s, 1H), 5.13 (d,J=14.0 Hz, 1H). MS (ESI): m/z 445.9 [M⁺+1]. HPLC: 98.55%.

Example 24

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(3-isopropylphenyl)pyridin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(24)

Compound 24 was prepared using the same conditions as compound 1 (0.020g). ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.75 (s, 1H), 7.99 (d,J=8.0 Hz, 1H), 7.79 (s, 1H), 7.65 (d, J=8.0 Hz, 1H), 7.45-7.33 (m, 5H),6.79-6.75 (m, 1H), 6.68-6.65 (m, 1H), 5.62 (d, J=14.5 Hz, 1H), 5.12 (d,J=14.5 Hz, 1H), 3.02-2.96 (m, 1H), 1.30 (d, J=7.0 Hz, 6H). MS (ESI): m/z472.1 [M⁺+1]. HPLC: 99.50%.

Example 25

2-(2,4-Difluorophenyl)-1-(5-(3,4-difluorophenyl)pyridin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(25)

Compound 25 was prepared using the same conditions as compound 1 (0.029g). ¹H NMR (500 MHz, CDCl₃): δ 8.75 (s, 1H), 8.67 (s, 1H), 7.94 (dd,J=8.0, 2.0 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H), 7.58 (br s, 1H), 7.42-7.36(m, 2H), 7.34-7.29 (m, 2H), 6.80-6.76 (m, 1H), 6.71-6.67 (m, 1H), 5.56(d, J=14.5 Hz, 1H), 5.17 (d, J=14.5 Hz, 1H). MS (ESI): m/z 466.0 [M⁺+1].HPLC: 98.94%.

Example 261-(5-(3-(Difluoromethoxy)phenyl)pyridin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(26)

Compound 26 was prepared using the same conditions as compound 1 (0.022g). ¹H NMR (500 MHz, CDCl₃): δ 8.79 (s, 1H), 8.77 (s, 1H), 7.98 (d,J=8.0 Hz, 1H), 7.67 (d, J=8.0 Hz, 1H), 7.57 (s, 1H), 7.51 (dd, J=8.0,2.0 Hz, 1H), 7.41-7.35 (m, 2H), 7.31 (s, 1H), 7.25-7.22 (m, 1H),6.79-6.74 (m, 1H), 6.69-6.62 (m, 1H), 6.59 (t, J=74.0 Hz, 1H), 5.58 (d,J=14.0 Hz, 1H), 5.17 (d, J=14.0 Hz, 1H). MS (ESI): m/z 496.0 [M⁺+1].HPLC: 92.30%.

Example 27

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-((trifluoromethyl)thio)phenyl)pyridin-2-yl)propan-2-ol(27)

Compound 27 was prepared using the same conditions as compound 1 (0.031g). ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.73 (s, 1H), 8.01 (d,J=8.0 Hz, 1H), 7.80 (d, J=8.5 Hz, 2H), 7.70 (d, J=8.0 Hz, 1H), 7.61 (d,J=8.5 Hz, 2H), 7.50 (br s, 1H), 7.42-7.37 (m, 1H), 6.80-6.76 (m, 1H),6.70-6.67 (m, 1H), 5.56 (d, J=14.5 Hz, 1H), 5.18 (d, J=14.5 Hz, 1H). MS(ESI): m/z 530.0 [M⁺+1]. HPLC: 96.42%.

Compounds 28-36 in Table 1 were prepared using the same conditions ascompound 1 from intermediate C and commercially available boronic acidsand azoles.

Compounds 37-42 in Table 1 were prepared using the same conditions ascompound 12 from intermediate E and commercially available aryl bromidesand azoles.

Compounds 43-45 in Table 1 were prepared using the same conditions ascompound 12 from intermediate E and aryl bromides that have beensynthesized via alkylation like intermediate F and commerciallyavailable azoles.

Example 46 and 47

2-(2,4-Difluorophenyl)-3-fluoro-1-(1H-tetrazol-1-yl)-3-(5-(4-(trifluoromethoxy)phenyl)-pyridin-2-yl)butan-2-ol(46 and 47)

To a stirred solution of 2,5-dibromopyridine (A; 30 g, 126.5 mmol) intoluene (1.5 L) was added n-BuLi (79 mL, 126 mmol; 1.6 M solution)dropwise at −78° C. under an inert atmosphere. After being stirred for40 min at −78° C., diethyl oxalate (20.6 mL, 126.5 mmol) was added tothe reaction mixture at −78° C. and stirring was continued for another20 min. After completion of the reaction (by TLC), the reaction mixturewas quenched with saturated NH₄Cl solution and extracted with EtOAc(2×1.0 L). The combined organic extracts were washed with water andbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to obtain the crude product. The crude material was purified bysilica gel column chromatography (eluent: 15-55% EtOAc/hexanes) toafford R (13 g, 50.37 mmol, 38%). ¹H NMR (200 MHz, CDCl₃): δ 8.81 (d,J=1.4 Hz, 1H), 8.17-7.98 (m, 2H), 4.48 (q, J=7.4 Hz, 2H), 1.41 (t, J=7.4Hz, 3H). MS (ESI): m/z 259 [M+1]⁺.

To a stirred solution of R (13 g, 50.3 mmol) in THF (150 mL) was addedmethyl magnesium chloride (CH₃MgCl; 15 mL, 50.3 mmol; 3 M solution inTHF) at −5° C. under an inert atmosphere. Stirring was continued foranother 2 h. Progress of the reaction was monitored by TLC. The reactionmixture was then quenched with saturated NH₄Cl solution and extractedwith EtOAc (2×200 mL). The combined organic extracts were washed withwater and brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude product. The crude material waspurified by silica gel column chromatography (eluent: 15-55%EtOAc/hexanes) to afford S (2.8 g, 10.76 mmol, 21%). ¹H NMR (200 MHz,CDCl₃): δ 8.61 (d, J=1.4 Hz, 1H), 7.84 (dd, J=8.0, 1.4 Hz, 1H), 7.49 (d,J=8.0 Hz, 1H), 4.92 (br s, 1H), 4.20 (q, J=7.4 Hz, 2H), 1.80 (s, 3H),1.22 (t, J=7.4 Hz, 3H).

To a stirred solution of S (2.8 g, 10.7 mmol) in CH₂C12 (50 mL) wasadded diethylaminosulfur trifluoride (DAST; 3.5 mL, 26.5 mmol) at 0° C.under an inert atmosphere, and the reaction mixture was stirred for 16 hat RT. Progress of the reaction was monitored by TLC. The reactionmixture was then quenched with ice-cold water and extracted with CH₂Cl₂(2×100 mL). The combined organic extracts were washed with water andbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude material was purified by silica gel columnchromatography (eluent: 15-55% EtOAc/hexanes) to afford T (2.1 g, 7.6mmol, 75%). ¹H NMR (200 MHz, CDCl₃): δ 8.62 (d, J=1.4 Hz, 1H), 7.85 (dd,J=8.0, 1.4 Hz, 1H), 7.50 (d, J=8.0 Hz, 1H), 4.23 (q, J=7.4 Hz, 2H), 1.95(d, J_(F,H)=24.0 Hz, 3H), 1.24 (t, J=7.4 Hz, 3H). MS (ESI): m/z 276[M]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.9 mL, 8.01 mmol)in diethyl ether (50 mL) was added dropwise n-BuLi (5 mL, 8.01 mmol; 1.6M solution) at −78° C. under an inert atmosphere. After being stirredfor 40 min at −78° C., a solution of T (2.1 g, 8.01 mmol) in diethylether (50 mL) was added dropwise to the reaction mixture at −78° C.Stirring was continued for another 20 min. After completion of thereaction (by TLC), the reaction mixture was quenched with saturatedNH₄Cl solution and extracted with EtOAc. The combined organic extractswere washed with water and brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to obtain the crude product. Thecrude material was purified by silica gel column chromatography (eluent:15-55% EtOAc/hexanes) to afford ketone U (2.15 g, 6.24 mmol, 77.9%). ¹HNMR (200 MHz, CDCl₃): δ 8.61 (d, J=1.6 Hz, 1H), 7.96 (dd, J=8.0, 1.6 Hz,1H), 7.67-7.62 (m, 1H), 7.48 (d, J=8.0 Hz, 1H), 6.98-6.67 (m, 2H), 1.98(d, J_(F,H)=24.0 Hz, 3H). MS (ESI): m/z 343.9 [M+1]⁺.

To a stirred solution of ketone U (2.1 g, 6.10 mmol) in acetonitrile(CH₃CN; 30 mL) were added iodotrimethylsilane (TMS-I; 1.47 g, 6.71 mmol)and potassium hydroxide (KOH; 683 mg, 12.20 mmol) at RT under an inertatmosphere. The resulting reaction mixture was heated to 70° C. andstirred for 1.5 h; progress of the reaction was monitored by TLC. Thereaction mixture was then diluted with EtOAc, stirred for 5 min andfiltered; the filtrate was concentrated under reduced pressure to obtainthe crude product. The crude material was purified by silica gel columnchromatography (eluent: 15-55% EtOAc/hexanes) to afford epoxide V (1.92g, 5.36 mmol, 88%) as a mixture of diastereomers. The product wasconfirmed by ¹H-NMR spectral analysis and was taken forward to the nextstep without any further purification.

To a stirred solution of epoxide V (1.0 g, 2.79 mmol) in THF (15 mL) andwater (5 mL) were added (4-(trifluoromethoxy)phenyl)boronic acid (575mg, 2.79 mmol), K₂CO₃ (770 mg, 5.58 mmol) and Pd(dppf)₂Cl₂ (102 mg,0.139 mmol) at RT under an inert atmosphere. The reaction mixture wasdegassed for 30 min by purging with argon. The reaction mixture was thenheated to 65° C. and stirred for 2 h; progress of the reaction wasmonitored by TLC. The reaction mixture was then cooled to RT, dilutedwith water and extracted with EtOAc (2×30 mL). The combined organiclayer was washed with water and brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to obtain the crude product. Thecrude compound was purified by silica gel column chromatography toafford W (0.9 g, 2.05 mmol, 79%) as a mixture of diastereomers. Theproduct was confirmed by ¹H-NMR and MS analyses and was taken forward tothe next step without any further purification. MS (ESI): m/z 440[M+1]⁺. To a stirred solution of W (900 mg, 2.05 mmol) in DMF (10 mL)was added 1H-tetrazole (215 mg, 3.07 mmol) followed by K₂CO₃ (283 mg,2.05 mmol) at RT under an inert atmosphere.

The resulting reaction mixture was heated to 65° C. and stirred for 48h; progress of the reaction was monitored by TLC. The reaction mixturewas then cooled to RT, diluted with water and extracted with EtOAc (2×50mL). The combined organic layers were washed with water and brine, driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude product. The crude material was purified by silica gel columnchromatography to afford the desired diastereomers 46 (110 mg, 0.21mmol, 11%) and 47 (120 mg, 0.23 mmol, 11.5%). 46: ¹H NMR (500 MHz,CDCl₃): δ 8.68 (s, 1H), 8.60 (s, 1H), 8.07 (d, J=8.5 Hz, 1H), 7.85-7.78(m, 3H), 7.63 (d, J=8.5 Hz, 2H), 7.38 (d, J=8.5 Hz, 2H), 6.90-6.87 (m,1H), 6.85-6.81 (m, 1H), 5.47 (d, J=14.5 Hz, 1H), 4.41 (d, J=14.5 Hz,1H), 1.50 (d, J_(F,H)=23.5 Hz, 3H). MS (ESI): m/z 510 [M+1]⁺. HPLC:99.73%. 47: ¹H NMR (500 MHz, CDCl₃): δ 8.73 (s, 1H), 8.66 (s, 1H), 7.76(dd, J=8.5, 2.0 Hz, 1H), 7.53 (d, J=8.5 Hz, 2H), 7.40 (br s, 1H),7.33-7.25 (m, 3H), 6.89-6.85 (m, 1H), 6.64-6.60 (m, 1H), 6.41-6.38 (m,1H), 5.76 (d, J=14.5 Hz, 1H), 5.00 (d, J=14.5 Hz, 1H), 1.98 (d,J_(F-H)=22.5 Hz, 3H). MS (ESI): m/z 510 [M+H]⁺. HPLC: 99.48%.

Compounds 48-57 in Table 1 were prepared using the same conditions ascompound 46 from intermediate V and commercially available boronic acidsand azoles.

Example 58

2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yldihydrogen phosphate (58)

To a suspension of 11(+) (200 mg, 0.38 mmol) and 1H-tetrazole (106 mg,1.51 mmol) in anhydrous CH₂Cl₂ (15 mL) was added a solution ofdibenzyl-N,N-diisopropylphosphoramidite (0.38 mL, 1.14 mmol) in CH₂C12(5 mL). After 48 h, the reaction mixture was cooled to −5° C., and asolution of 3-chloroperoxybenzoic acid (m-CPBA; 195 mg, 1.14 mmol) inCH₂C12 (5 mL) was added slowly. After 1 h, the mixture was diluted withCH₂C12 (30 mL), washed with 5% aqueous Na₂S₂O₅ (2×30 mL), 10% aqueousNaHCO₃ (2×30 mL) and brine (30 mL) and then dried over anhydrous Na₂SO₄.The organic solvent was evaporated in vacuo, and the obtained crudematerial was purified by preparative HPLC to afford compound X (125 mg,0.16 mmol, 42%) as colorless semisolid. ¹H NMR (500 MHz, CDCl₃): δ 9.12(s, 1H), 8.82 (s, 1H), 7.75 (d, J=8.5 Hz, 1H), 7.55 (d, J=9.0 Hz, 2H),7.37 (m, 5H), 7.31-7.18 (m, 7H), 7.08 (d, J=9.0 Hz, 2H), 6.70-6.58 (m,2H), 6.24 (d, J=16.0 Hz, 1H), 5.97 (d, J=16.0 Hz, 1H), 5.25-5.18 (m,2H), 4.97-4.84 (m, 2H), 4.42 (q, J=8.5 Hz, 2H). ³¹P NMR (500 MHz,CDCl₃): δ −14.29 (s); HPLC: 99%. MS(ESI): m/z 788 [M+H]⁺.

A mixture of compound X (250 mg, 0.32 mmol) and 10% Pd/C (50 mg) in EtOH(10 mL) was stirred under hydrogen (1 atm) for 3 h. The catalyst wasremoved by filtration over a pad of Celite®, and the filter cake waswashed with EtOH (5 mL) and EtOAc (5 mL). The combined filtrates wereevaporated in vacuo, and the residue was triturated with n-pentane togive 58 (140 mg, 0.23 mmol, 72%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆): δ 9.39 (s, 1H), 8.90 (s, 1H), 8.19 (d, J=8.0 Hz, 1H), 7.79 (d,J=9.0 Hz, 2H), 7.52 (d, J=8.0 Hz, 1H), 7.35-7.30 (m, 1H), 7.21-7.14 (m,3H), 6.98-6.95 (m, 1H), 6.18 (d, J=15.5 Hz, 1H), 5.79 (d, J=15.5 Hz,1H), 4.85 (q, J=9.0 Hz, 2H). ³¹P NMR (500 MHz, DMSO-d₆): δ −6.57 (s).HPLC: 99%. MS(ESI): m/z 608 [M+1-1]⁺.

HPLC Method A Specifications

Column: Aquity BEH C-18 (50×2.1 mm, 1.7μ)

Mobile Phase: A) Acetonitrile; B) 0.025% aqueous (aq) trifluoroaceticacid (TFA)

Flow Rate: 0.50 mL/min

Time (min)/% B: 0.01/90, 0.5/90, 3/10, 6/10

HPLC Method O Specifications:

Column: Zorbax Phenyl Hexyl (50×4.6 mm, 1.80

Mobile Phase: A) Acetonitrile; B) 0.1% as TFA

Flow Rate: 1.00 mL/min

Time (min)/% B: 0.01/50, 1/50, 4/10, 10/10

TABLE 1 Structures for Example Compounds Example Number Structure 28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

TABLE 2 Analytical Data for Example Compounds in Table 1 HPLC RetentionExample HPLC Time ESIMS Number Method (min) (M + H) 28 A  2.9 501.9 29 A2.77 482.0 30 A 2.26 446.1 31 A 2.76 498.0 32 A 2.51 475.1 33 A  2.3526.1 34 A 2.95 531.0 35 A 2.54 430.0 36 A 2.46 515.0 37 A 2.52 541.0 38A 2.64 496.0 39 A 2.82 513.0 40 A 2.74 512.1 41 A 2.75 512.0 42 A 2.05487.1 43 O 3.81 44 A  2.8 542.0 45 A  2.8 544.1 46 A 3.14 510.2 47 A3.01 510.2 48 A 3.02 444.5 49 A 2.81 444.6 50 A  2.9 444.5 51 A 2.73444.5 52 A 3.07 494.0 53 A 2.88 494.2 54 A 3.06 460.2 55 A 2.92 460.1 56A 3.04 524.3 57 A 2.85 524.1 58 A 2.35 608  

Example 59: Metalloenzyme Activity

A. Minimum Inhibitory Concentration (MIC)

Compounds were assessed for their ability to inhibit the growth ofcommon strains of fungus, C. albicans using a standardized procedure(CLSI M27-A2).

Stock solutions of the test compounds and standards were prepared inDMSO at 1,600 μg/mL (C. albicans). Eleven, serial, one-half dilutions ofcompounds were prepared in 96-well plates in RPMI+MOPS. The assayconcentration ranges were 8-0.001 μg/mL (C. albicans). Cell suspensionsof C. albicans were prepared and added to each well at concentrations ofapproximately 3.7×10³ colony-forming-units per milliliter (cfu/mL). Alltesting was in duplicate. The inoculated plates were incubated forapproximately 48 h at 35±1° C. At the completion of incubation the wellsof each plate were evaluated visually for the presence of fungal growth.

For fluconazole and the test compounds, the MIC was the concentration atwhich growth was significantly reduced (about 50% reduction). Forvoriconazole the MIC was the concentration which reduced C. albicansgrowth by 50% (per CLSI, M27-A2). For QC purposes C. krusei isolate ATCC6258 (4.0×10³ cfu/mL) was included in the VOR assay. This isolate didnot exhibit trailing growth against voriconazole, therefore the MIC wasthe concentration at which growth was completely inhibited.

Example 60: Metalloenzyme Selectivity

A. Inhibition of Liver Cytochrome P450 Enzymes

Solutions of each test compound were separately prepared atconcentrations of 20000, 6000, 2000, 600, 200, and 60 μM by serialdilution with DMSO:MeCN (50:50 v/v). The individual test compoundsolutions were then diluted 20-fold with DMSO:MeCN:deionized water(5:5:180 v/v/v) to concentrations of 1000, 300, 100, 30, 10, and 3 μM.Mixtures of isozyme inhibitors (sulfaphenazole, tranylcypromine, andketoconazole as specific inhibitors of isozymes 2C9, 2C19, and 3A4,respectively) were prepared containing each inhibitor at concentrationsof 6000, 2000, 600, 200, 60, 20, 6, and 2 μM by serial dilution withDMSO:ACN (50:50 v/v). The mixed inhibitor solutions were then diluted20-fold with DMSO:MeCN:deionized water (5:5:180 v/v/v) to concentrationsof 300, 100, 30, 10, 3, 1, 0.3, and 0.1 μM. The percent of organicsolvent attributable to the test compound or inhibitor mixture in thefinal reaction mixture was 2% v/v.

Pooled human liver microsome suspension (20 mg/mL) was diluted withphosphate buffer to obtain a 5 mg/mL suspension. A solution of NADPH wasprepared in phosphate buffer at a concentration of 5 mM. Separate stocksolutions of each substrate were prepared in DMSO:MeCN (50:50 v/v),mixed, and diluted in phosphate buffer to obtain a single solutioncontaining each substrate at five times its experimentally determinedK_(m) concentration. The percent of organic solvent attributable tosubstrate mixture in the final reaction mixture was 1% v/v.

Substrate solution and microsome suspension were combined in a 1:1volume ratio, mixed, and distributed to reaction wells of a PCR plate.Individual test compound or combined inhibitor solutions at eachconcentration were added to the wells and mixed by repetitiveaspirate-dispense cycles. For active controls, blank phosphate buffersolution was added in place of test compound solution. Reaction mixtureswere allowed to equilibrate at 37° C. for approximately two minutesbefore adding NADPH solution to initiate reaction, followed by pipettemixing of reaction mixture. Ten minutes after addition of NADPH, thereaction mixtures were quenched with cold acetonitrile. The samples weremixed by orbital shaking for approximately one minute and centrifuged at2900 RCF for ten minutes. A portion of the supernatant was analyzed bygradient reverse-phase HPLC with detection by electrospray ionizationtriple quadrupole mass spectrometry in the positive ion mode.

Data was fitted to sigmoid dose-response curves and the inhibitorypotency of each test compound was determined as its IC₅₀ value.

Results

Candida CYP2C9 CYP2C19 CYP3A4 Example MIC* IC50 IC50 IC50 1 ≤0.016 >6035 16 2 ≤0.016 >60 >60 >60 Fluconazole 0.5  29 8.2 8.0 Voriconazole0.016  14 15 13 *Candida albicans MIC (median inhibitory concentration)values expressed in ug/mL; CYP IC5Os are in uM.Compound examples 3-28 exhibit Candida MICs in the range as indicatedbelow.In each case of Table 3 the rating scale is as follows:

MIC (μg/mL Rating ≤0.5 A >0.5 − 1.5 B >1.5 − 4 C >4 D Not tested E

TABLE 3 MIC Data for Compounds in Table 1 Candida Example MIC NumberRating  1 A  2 A  3 A  4 A  5 A  6 A  7 A  8 A  9 A 10 A 11 A 12 A 13 E14 E 15 A 16 A 17 E 18 A 19 E 20 D 21 C 22 C 23 A 24 A 25 A 26 A 27 A 28A 29 A 30 A 31 A 32 A 33 A 34 A 35 A 36 A 37 A 38 A 39 C 40 A 41 A 42 A43 A 44 A 45 A 46 B 47 A 48 C 49 C 50 C 51 A 52 C 53 B 54 C 55 A 56 C 57A 58 A

INCORPORATION BY REFERENCE

The contents of all references (including literature references, issuedpatents, published patent applications, and co-pending patentapplications) cited throughout this application are hereby expresslyincorporated herein in their entireties by reference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents of the specificembodiments of the invention described herein. Such equivalents areintended with be encompassed by the following claims.

What is claimed:
 1. A method of treating a subject suffering from orsusceptible to a disorder or disease, wherein the subject has beenidentified as in need of treatment for the disorder or disease,comprising administering to said subject in need thereof, an effectiveamount of a compound of Formula I,

or salt thereof, wherein: MBG is tetrazolyl; R₁ is halo; R₂ is halo; R₃is independently halo, haloalkoxy, haloalkoxyalkyl, or hydroxyl; R₄ isaryl, optionally substituted with 0, 1, 2 or 3 independent R₃; R₅ is Hor —P(O)(OH)₂; R₆ is H or halo; and n is 0, 1, 2 or
 3. 2. The method ofclaim 1, wherein the disorder or disease is associated with one or moreof the following pathogenic fungi: Absidia corymbifera, Ajellornycesdermatitidis, Arthroderma benhamiae, Arthroderma flavum, Arthrodermagypseum, Arthroderma incurvaturn, Arthroderma otae, Arthrodermavanbreuseghemii, Aspergillus flavus, Aspergillus fumigates, Aspergillusniger, Blastomyces dermatitidis, Candida albicans, Candida glabrata,Candida guilliermondii, Candida krusei, Candida parapsilosis, Candidatropicalis, Candida pelliculosa, Cladophialophora carrionii,Coccidioides immitis, Cryptococcus neoformans, Cunninghamella sp.,Epidermophyton floccosum, Exophiala dermatitidis, Filobasidiellaneoformans, Fonsecaea pedrosoi, Fusarium solani, Geotrichum candidum,Histoplasma capsulatum, Hortaea werneckii, Issatschenkia orientalis,Madurella grisae, Malassezia fur, Malassezia globosa, Malassezia obtusa,Malassezia pachydermatis, Malassezia restricta, Malassezia slooffiae,Malassezia sympodialis, Microsporum canis, Microsporum fulvum,Microsporum gypseum, Mucor circinelloides, Nectria haematococca,Paecilomyces variotii, Paracoccidioides brasiliensis, Penicilliummarneffei, Pichia anomala, Pichia guilliermondii, Pneumocystis carinii,Pseudallescheria boydii, Rhizopus oryzae, Rhodotorula rubra,Scedosporium apiospernium, Schizophyllum commune, Sporothrix schenckii,Trichophyton mentagrophytes, Trichophyton rubrum, Trichophytonverrucosum, Trichophyton violaceum, Trichosporon asahii, Trichosporoncutaneum, Trichosporon inkin, Trichosporon mucoides.
 3. The method ofclaim 2, wherein the compound is2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol.4. The method of claim 2, wherein the compound is(+)-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol.5. The method of claim 1, wherein the disorder or disease isAspergillosis, Blastomycosis, Candidiasis, Chromomycosis,Coccidioidomycosis, Cryptococcosis, Dermatophytoses, Histoplasmosis,Keratomycosis, Lobomycosis, Malassezia infection, Mucormycosis,Paracoccidioidomycosis, Penicillium marneffei infection,Phaeohyphomycosis, Pneumocyctis pneumonia, or Rhinosporidiosis.
 6. Themethod of claim 5, wherein the compound is22-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol.7. The method of claim 5, wherein the compound is(+)-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol.8. The method of claim 1, wherein R₁ is fluoro; R₂ is fluoro; R₃ ishaloalkoxy or haloalkoxyalkyl; R₄ is phenyl substituted with 1, 2, or 3fluoro; R₅ is hydrogen; R₆ is hydrogen; and n is
 1. 9. The method ofclaim 8, herein R₂ is fluoro.
 10. The method of claim 9, wherein R₁ andR₂ are fluoro.
 11. The method of claim 1, wherein R₁ is fluoro.
 12. Themethod of claim 1, wherein wherein R₃ is haloalkoxyalkyl or haloalkoxy,and n is
 1. 13. The method of claim 12, wherein wherein R₃ is2,2,2-trifluoroethoxy, and n is
 1. 14. The method of claim 1, whereinwherein R₄ is phenyl optionally substituted with 0, 1, 2, or 3independent halo.
 15. The method of claim 14, wherein wherein R₄ isphenyl optionally substituted with 2 halo.
 16. The method of claim 15,wherein wherein R₄ is phenyl substituted with 2 fluoro.
 17. The methodof claim 1, wherein wherein R₅ is hydrogen.
 18. The method of claim 1,wherein wherein R₁ and R₂ are fluoro; R₃ is 2,2,2-trifluoroethoxy and nis 1; R₄ is difluorophenyl; and R₅ and R₆ are hydrogen.
 19. The methodof claim 1, wherein the compound is22-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-olor2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yldihydrogen phosphate.
 20. The method of claim 1, wherein the compound is22-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol.21. The method of claim 1, wherein the compound is(+)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol.